AU2018310983B2

AU2018310983B2 - Optimized peptides for targeting human nerves and their use in image guided surgery, diagnostics and therapeutic delivery

Info

Publication number: AU2018310983B2
Application number: AU2018310983A
Authority: AU
Inventors: Stephen Adams; Dina HINGORANI; Quyen T. Nguyen; Roger Y. Tsien; Mike A. WHITNEY
Original assignee: University of California Berkeley; University of California San Diego UCSD
Current assignee: University of California San Diego UCSD
Priority date: 2017-08-02
Filing date: 2018-08-02
Publication date: 2025-01-23
Anticipated expiration: 2038-08-02
Also published as: EP3661539A4; EP3661539B1; EP4484952A3; IL309994B1; US11021517B2; CA3071835A1; US20210363186A1; CN111246873A; WO2019028281A3; US11299515B2; WO2019028281A2; EP3661539A2; JP7282087B2; JP2023103369A; US20200239522A1; IL309994A; US20240124523A1; EP4484952A2; US20210087229A1; KR102929599B1

Abstract

The present invention provides methods for guiding preservation of human neurons or human nerves during surgery by administering a fluorescently-labeled peptide that specifically binds to the human neurons or human nerves. The invention further provides human neuron or nerve targeting molecules comprising fluorescently-labeled peptides that specifically bind to human neurons or human nerves and compositions thereof.

Description

OPTIMIZED PEPTIDES FOR TARGETING HUMAN NERVES AND THEIR USE IN IMAGE GUIDED SURGERY, DIAGNOSTICS AND THERAPEUTIC DELIVERY CROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims priority to U.S. Provisional Application No. 62/659,612, filed April 18, 2018, and U.S. Provisional Application No. 62/540,510, filed August 02, 2017, both of which are

incorporated herein by reference in their entireties.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] This invention was made with Government support under EB008122 and EB014929 awarded by the National Institutes of Health. The Government has certain rights in the invention

BACKGROUND OF THE INVENTION

[0003] Preservation of human neurons and human nerves is one of the most important goals of any surgical procedure, because accidental transection of neuron or nerves leads to significant

morbidity. Nerves are typically identified by their elongated whitish appearance and relationship to

nearby structures or by electrophysiological studies. However, in instances such as trauma, tumor

involvement, inflammation, or infection, nerve identification using these criteria can be difficult.

Therefore, there is a need for methods of reliably and conclusively identifying neuron or nerves

which overcome the deficiencies in the art.

[0004] Neuron or nerve identification prior to direct exposure during surgery or confirmation of neuron or nerve identity in instances of uncertainty following direct exposure is accomplished by

electromyographic (EMG) monitoring. This technique, however, has the disadvantage of not

providing visual feedback to the operating surgeon. Thus, even if a nerve has been identified in one

location, either through accidental or purposeful stimulation, there is no visual guidance to the

operating surgeon as to how far away from the stimulation site the nerve lies or the direction of travel the nerve takes away from the stimulation site. Furthermore, EMG only traces motor pathways, not sensory fibers. EMG fails if neuron or nerve conduction or neuromuscular transmission is temporarily blocked anywhere distal to the recording site. Such blockade easily occurs due to neuron or nerve compression, trauma, local anesthetics, or neuromuscular blockers.

[0005] Neuron or nerve labeling primarily depend on retrograde or anterograde tracing of individually identified axonal tracts via the use of fluorescent dyes. However, methods of labeling

neuron or nerves by locally applied fluorescent tracers have several disadvantages. First, this

technique can label only one neuron or nerve fiber tract at a time, depending on where the dye has been injected. Second, this technique results in only limited labeling of fluorescent dyes along the

axonal tracts, because retrograde axonal tracers typically accumulate in the neural cell body. Third,

retrograde transport is relatively slow (on the order of millimeters per day) and therefore takes a

long time to label human neuron or nerves, which are often longer than a meter, such as in the case

of the sciatic neuron or nerve and its arborizations. Fourth, the application of fluorescent dyes to

innervation targets such as direct intramuscular injections to label motor neuron or nerves is

typically messy with a variable amount of the tracer dye remaining at the injection site. As dissection

of neuron or nerves depends on accurate visualization of adjacent structures prior to encountering

them, a surgical site that is contaminated with fluorescent dyes would not be desirable. Finally, the

direct injection of the fluorescent dye itself may be damaging to the target organs or neuron or

nerve of interest, either by mechanical damage or by the very high local concentration of dye and

vehicle at the injection site.

[0006] There has been a need in the art to identify peptides capable of binding to human nerves and neurons, in order to facilitate surgical procedures and human nerve protection.

[0007] Nerve-homing peptides sequences were previously identified by their ability to bind mouse nerves for laboratory research. However, the peptide sequences described in the present

application were identified by their ability to bind human nerves, following systemic intravenous

injection into human patients and as such these peptides meet the need of being able to more

specifically and effectively bind to human nerves compared to previous sequences. The present

invention provides peptide sequences that selective bind to human nerves and/or neurons, as well

as methods of using those sequences in surgical procedures, for example to preserve nerves and/or

to avoid nerve damage during such procedures.

[0007a] Any discussion of the prior art throughout the specification should in no way be considered as an admission that such prior art is widely known or forms part of common general knowledge in the field.

BRIEF SUMMARY OF THE INVENTION

[0007b] According to a first aspect, the present invention provides a human neuron or nerve targeting molecule that specifically binds to a human neuron or nerve, or component of either, wherein said targeting molecule comprises a peptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9), Ac PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13), Ac SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS (HNP401-N-8; SEQ ID NO:24), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), SGQVPWEEPYYVV (HNP401-C 4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),

SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), and SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124).

[0007c] According to a second aspect, the present invention provides a multidomain neuron or nerve targeting molecule comprising two or more neuron or nerve targeting peptides, wherein the two or more neuron or nerve targeting peptides bind to a human neuron or nerve, or component of either, wherein the first peptide is selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9), Ac PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13), Ac SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS (HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), SGQVPWEEPYYVV (HNP401-C 4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122), SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), and 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

[0007d] According to a third aspect, the present invention provides a method of identifying a human neuron or nerve comprising contacting the human neuron or nerve with a targeting molecule according to the first aspect and further comprising a cargo selected from the group consisting of drug, fluorescent moiety, and photosensitizing agent, wherein the cargo comprises the fluorescent moiety.

[0007e] According to a fourth aspect, the present invention provides a method of delivering a drug to a human neuron or nerve comprising contacting the human neuron or nerve with a human neuron or nerve targeting molecule according to the first aspect and further comprising a cargo selected from the group consisting of a drug, fluorescent moiety, and photosensitizing agent, wherein the cargo comprises the drug.

[0007f] According to a fifth aspect, the present invention provides a method of delivering a photosensitizing agent to a human neuron or nerve comprising contacting the human neuron or nerve with a human neuron or nerve targeting molecule according to the first aspect and further comprising a cargo selected from the group consisting of a drug, fluorescent moiety, and photosensitizing agent, wherein the cargo comprises the photosensitizing agent.

[0007g] According to a sixth aspect, the present invention provides a pharmaceutical composition comprising: (a) the human neuron or nerve targeting molecule of the first aspect or the multidomain neuron or nerve targeting molecule of the second aspect, and (b) a pharmaceutically acceptable excipient

[0007h] Unless the context clearly requires otherwise, throughout the description and the claims, the words "comprise", "comprising", and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of "including, but not limited to".

[0008] Disclosed herein, in certain embodiments, are targeting molecules comprising a peptide that specifically binds to a human neuron, human nerve, or component of either. In some embodiments, the peptide is selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac-

DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9), Ac PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13), Ac SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSS (HNP401-N-2 with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS (HNP401-N-8; SEQ ID NO:24), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), SGQVPWEEPYYVV (HNP401-C 4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122), SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124),

[0009] 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104), PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122), SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), and/or combinations thereof.

[0010] In some embodiments, the human neuron or nerve targeting molecule that specifically binds to a human neuron or nerve, or component of either, wherein said targeting molecule comprises a

peptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),

WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3),

DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC

linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac

DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC

(HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker;

SEQ ID NO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9), Ac-PYYVVKKSSGGC

(HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC

linker; SEQ ID NO:11), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac

SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20);

QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;

SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS (HNP401-N-8; SEQ ID

NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28),

PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6

with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120),

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

(HNP401-C-4 with GG linker; SEQ ID NO:122), SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), 5FAM

QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104), PWEEPYYVVKKSSGG

(HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID

NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120), SGQVPWEEPYYVVKKGG

(HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker;

SEQ ID NO:122), SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG

(HNP401-C-8 with GG linker; SEQ ID NO:124), and/or combinations thereof.

[0011] In some embodiments the targeting molecule comprises a peptide selected from the group consisting: of SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ

ID NO:2), and DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

[0012] In some embodiments, the targeting molecule comprises a peptide selected from the group consisting of SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401

C-2 with GGC linker; SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ

ID NO:7), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG (HNP401-N-2

with GG linker; SEQ ID NO:21), and SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25).

[0013] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1).

[0014] In some embodiments, the targeting molecule comprises a peptide WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2).

[0015] In some embodiments, the targeting molecule comprises the peptide DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

[0016] In some embodiments, the targeting molecule comprises the peptide Ac SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4).

[0017] In some embodiments, the targeting molecule comprises the peptide Ac WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5).

[0018] In some embodiments, the targeting molecule comprises the peptide Ac DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6).

[0019] In some embodiments, the targeting molecule comprises the peptide Ac QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7).

[0020] In some embodiments, the targeting molecule comprises the peptide Ac PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8).

[0021] In some embodiments, the targeting molecule comprises the peptide Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9).

[0022] In some embodiments, the targeting molecule comprises the peptide Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10).

[0023] In some embodiments, the targeting molecule comprises the peptide Ac SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11).

[0024] In some embodiments, the targeting molecule comprises the peptide Ac SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12).

[0025] In some embodiments, the targeting molecule comprises the peptide Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13).

[0026] In some embodiments, the targeting molecule comprises the peptide Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14).

[0027] In some embodiments, the targeting molecule comprises the peptide QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20).

[0028] In some embodiments, the targeting molecule comprises the peptide QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).

[0029] In some embodiments, the targeting molecule comprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22).

[0030] In some embodiments, the targeting molecule comprises the peptide EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23).

[0031] In some embodiments, the targeting molecule comprises the peptide PYYVVKKSS (HNP401 N-8; SEQ ID NO:24).

[0032] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25).

[0033] In some embodiments, the targeting molecule comprises the SGQVPWEEPYYVV (HNP401-C 4; SEQ ID NO:26).

[0034] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27).

[0035] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEP (HNP401 C-8; SEQ ID NO:28).

[0036] In some embodiments, the targeting molecule comprises the peptide PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118).

[0037] In some embodiments, the targeting molecule comprises the peptide EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119).

[0038] In some embodiments, the targeting molecule comprises the peptide PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120).

[0039] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121).

[0040] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122).

[0041] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123).

[0042] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124).

[0043] In some embodiments, the targeting molecule comprises the peptide 5FAM QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

[0044] In some embodiments, the human neuron or nerve targeting molecule further comprises a cargo. In some embodiments, the cargo is a drug, a fluorescent moiety, a photosensitizing agent, or a combination thereof.

[0045] In some embodiments, the human neuron or nerve targeting molecule further comprises a drug.

[0046] In some embodiments, the human neuron or nerve targeting molecule further comprises a drug selected from the group consisting of: an antihistamine, a GABA receptor modulator, a neurotransmitter reuptake inhibitor, a local anesthetic, an anticholinergic, a sodium channel blocker, a calcium channel blocker, a thyrotropin-releasing hormone, ay-secretase inhibitor, an AMPA receptor agonist or antagonist, an NMDA receptor agonist or antagonist, an mGlu receptor agonist or antagonist, a growth factor, an antiemetic agent, a corticosteroid; a cytotoxic agent; an antioxidant, an iron chelator, a mitochondrial modulator, a sirtuin modulator, a nitric oxide (NO) and/or nitric oxide synthase (NOS) modulator, a potassium channel agonist or antagonist, a purigenic receptor agonist or antagonist, and/or combinations thereof.

[0047] In some embodiments, the human neuron or nerve targeting molecule further comprises a drug selected from the group consisting of: benzocaine; carticaine; cinchocaine; cyclomethycaine; lidocaine; prilocaine; propxycaine; proparacaine; tetracaine; tocainide; and trimecaine;

methotrexate; cyclophosphamide; thalidomide; paclitaxel; pemetrexed; pentostatin; pipobroman;

pixantrone; plicamycin; platonin; procarbazine; raltitrexed; rebeccamycin; rubitecan; SN-38;

salinosporamide A; satraplatin; streptozotocin; swainsonine; tariquidar; taxane; tegafur-uracil;

temozolomide; testolactone; thioTEPA; tioguanine; topotecan; trabectedin; tretinoin; triplatin

tetranitrate; tris(2- chloroethyl)amine; troxacitabine; uracil mustard; valrubicin; vinblastine;

vincristine; vinorelbine; vorinostat; zosuquidar; carbamazepine; oxcarbazepine; phenytein; valproic

acid; sodium valproate; cinnarizine; flunarizine; nimodipine; brain-derived neurotrophic factor

(BDNF); ciliary neurotrophic factor (CNTF); glial cell-line derived neurotrophic factor (GDNF);

neurotrophin-3; neurotrophin-4; fibroblast growth factor (FGF) receptor; insulin-like growth factor

(IGF); and/or combinations thereof.

[0048] In some embodiments, the human neuron or nerve targeting molecule further comprises a fluorescent moiety.

[0049] In some embodiments, the human neuron or nerve targeting molecule further comprises a fluorescent moiety selected from the group consisting of: a fluorescent protein, a fluorescent

peptide, a fluorescent dye, and/or combinations thereof.

[0050] In some embodiments, the human neuron or nerve targeting molecule further comprises a fluorescent moiety selected from the group consisting: of a xanthene; a bimane; a coumarin; an

aromatic amines; a benzofuran; a fluorescent cyanine; a carbazole; a dicyanomethylene pyrane;

polymethine; oxabenzanthrane; pyrylium; carbostyl; perylene; acridone; quinacridone; rubrene;

anthracene; coronene; phenanthrecene; pyrene; butadiene; stilbene; porphyrin; pthalocyanine;

lanthanide metal chelate complexes; rare-earth metal chelate complexes; derivatives thereof,

and/or combinations thereof.

[0051] In some embodiments, the human neuron or nerve targeting molecule further comprises a fluorescent moiety selected from the group consisting: of 5-carboxyfluorescein; fluorescein-5

isothiocyanate; 6-carboxyfluorescein; 5(6)-carboxyfluorescein; tetramethylrhodamine-6

isothiocyanate; 5-carboxytetramethylrhodamine; 5-carboxy rhodol derivatives; tetramethyl and

tetraethyl rhodamine; diphenyldimethyl and diphenyldiethyl rhodamine; dinaphthyl rhodamine;

rhodamine 101 sulfonyl chloride; Cy3, Cy3B, Cy3.5, Cy5, Cy5.5, Cy 7, indocyanine green, IR800CW,

cyan fluorescent protein (CFP), EGFP, 6-FAM, FAM, fluorescein, 5,6-dicarboxyfluorescein, 5-(and 6)

sulfofluorescein, sulfonefluorescein, succinyl fluorescein, 5-(and 6)-carboxy SNARF-1,

carboxyfluorescein sulfonate, carboxyfluorescein zwitterion, carboxyfluorescein quaternary

ammonium, carboxyfluorescein phosphonate, carboxyfluorescein GABA, carboxyfluorescein-cys-Cy5,

5'(6')-carboxyfluorescein, fluorescein glutathione, and/or combinations thereof.

[0052] In some embodiments, the human neuron or nerve targeting molecule further comprises a photosensitizing agent.

[0053] In some embodiments, the human neuron or nerve targeting molecule further comprises a photosensitizing agent selected from the group consisting of: a porphyrin, chlorin, and dye.

[0054] In some embodiments, the human neuron or nerve targeting molecule further comprises a photosensitizing agent selected from the group consisting of: porphyrin, protoporfin IX, purlytin,

verteporfin, HPPH, temoporfin, methylene blue, photofrin, protofrin, hematoporphyrin, Talaporfin, benzopophyrin derivative monoacid, 5-aminileuvolinic acid, Lutetium texaphyrin,

metallophthalocyanine, metallo-naphthocyaninesulfobenzo-porphyrazine, metallo

naphthalocyanines\, zinc tetrasulfophthalocyanine, bacteriochlorins, metallochlorins, chlorine

derivative, Tetra(m-hydroxyphenyl)chlorin (mTHPC), pheophorbide, dibromofluorescein (DBF),

IR700DX, naphthalocyanine, porphyrin derivative, and/or combinations thereof.

[0055] In some embodiments, provided is a method of identifying a human neuron or nerve comprising contacting the human neuron or nerve with a targeting molecule comprising (a) a

peptide that specifically binds to the human neuron or nerve, or component of either, and (b) a

fluorescent moiety, wherein said targeting molecule comprises a peptide selected from the group

consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ

ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID

NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac

WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC

(HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;

SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac

EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9), Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID

NO:11), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20);

NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), SGQVPWEEPYYVV (HNP401-C-4; SEQ ID

NO:26), SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28),

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), 5FAM QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104), and/or combinations

thereof.

[0056] In some embodiments, the human neuron or nerve targeting molecule comprises a peptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1),

WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

[0057] In some embodiments, the human neuron or nerve targeting molecule comprises a peptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), Ac

SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC

(HNP401-N-2 with GGC linker; SEQ ID NO:7), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20),

QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21), and SGQVPWEEPYYVVKK

(HNP401-C-2; SEQ ID NO:25).

[0058] In some embodiments, the targeting molecule comprises the peptic SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1).

[0059] In some embodiments, the targeting molecule comprises the peptide WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2).

[0060] In some embodiments, the targeting molecule comprises the peptide DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

[0061] In some embodiments, the targeting molecule comprises the peptide Ac SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4).

[0062] In some embodiments, the targeting molecule comprises the peptide Ac WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5).

[0063] In some embodiments, the targeting molecule comprises the peptide Ac DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6).

[0064] In some embodiments, the targeting molecule comprises the peptide Ac QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7).

[0065] In some embodiments, the targeting molecule comprises the peptide Ac PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8).

[0066] In some embodiments, the targeting molecule comprises the peptide Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9).

[0067] In some embodiments, the targeting molecule comprises the peptide Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10).

[0068] In some embodiments, the targeting molecule comprises the peptide Ac SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11).

[0069] In some embodiments, the targeting molecule comprises the peptide Ac SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12).

[0070] In some embodiments, the targeting molecule comprises the peptide Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13).

[0071] In some embodiments, the targeting molecule comprises the peptide Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14).

[0072] In some embodiments, the targeting molecule comprises the peptide DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16).

[0073] In some embodiments, the targeting molecule comprises the peptide QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20).

[0074] In some embodiments, the targeting molecule comprises the peptide QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).

[0075] In some embodiments, the targeting molecule comprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22).

[0076] In some embodiments, the targeting molecule comprises the peptide EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23).

[0077] In some embodiments, the targeting molecule comprises the peptide PYYVVKKSS (HNP401 N-8; SEQ ID NO:24).

[0078] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25).

[0079] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26).

[0080] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27).

[0081] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEP (HNP401 C-8; SEQ ID NO:28).

[0082] In some embodiments, the targeting molecule comprises the peptide PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118).

[0083] In some embodiments, the targeting molecule comprises the peptide EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119).

[0084] In some embodiments, the targeting molecule comprises the peptide PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120).

[0085] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121).

[0086] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122).

[0087] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123).

[0088] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124).

[0089] In some embodiments, the fluorescent moiety is selected from the group consisting of: a fluorescent protein, a fluorescent peptide, a fluorescent dye, and/or combinations thereof.

[0090] In some embodiments, the targeting molecule comprises the peptide 5FAM QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

[0091] In some embodiments, the fluorescent moiety is selected from the group consisting: of a xanthene; a bimane; a coumarin; an aromatic amine; a benzofuran; a fluorescent cyanine; a carbazole; a dicyanomethylene pyrane; polymethine; oxabenzanthrane; pyrylium; carbostyl; perylene; acridone; quinacridone; rubrene; anthracene; coronene; phenanthrecene; pyrene; butadiene; stilbene; porphyrin; pthalocyanine; lanthanide metal chelate complexes; rare-earth metal chelate complexes; derivatives thereof, and/or combinations thereof.

[0092] In some embodiments, the fluorescent moiety is selected from the group consisting of: 5 carboxyfluorescein (5-FAM); fluorescein-5-isothiocyanate; 6-carboxyfluorescein (6-FAM); 5(6) carboxyfluorescein; tetramethylrhodamine-6-isothiocyanate; 5-carboxytetramethylrhodamine; 5 carboxy rhodol derivatives; tetramethyl and tetraethyl rhodamine; diphenyldimethyl and diphenyldiethyl rhodamine; dinaphthyl rhodamine; rhodamine 101 sulfonyl chloride; Cy3, Cy3B, Cy3.5, Cy5, Cy5.5, Cy 7, indocyanine green, IR800CW, cyan fluorescent protein (CFP), EGFP, 6-FAM, FAM, fluorescein, 5,6-dicarboxyfluorescein, 5-(and 6)-sulfofluorescein, sulfonefluorescein, succinyl fluorescein, 5-(and 6)-carboxy SNARF-1, carboxyfluorescein sulfonate, carboxyfluorescein zwitterion, carboxyfluorescein quaternary ammonium, carboxyfluorescein phosphonate, carboxyfluorescein

GABA, carboxyfluorescein-cys-Cy5, 5'(6')-carboxyfluorescein, fluorescein glutathione, and/or

combinations thereof.

[0093] In some embodiments, provided is a method of delivering a drug to a human neuron or nerve comprising contacting the human neuron or nerve with a human neuron or nerve targeting

molecule comprising (a) a peptide that specifically binds to the neuron or nerve, or component of

either, and (b) a drug, wherein said targeting molecule comprises a peptide selected from the group

consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID

NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac

WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC

SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac

EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9), Ac-PYYVVKKSSGGC (HNP401-N-8

with GGC linker; SEQ ID NO:10), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID

NO:11), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20);

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124),

[0094] 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104), and/or combinations thereof.

[0095] In some embodiments, the drug is selected from the group consisting of: an antihistamine, a GABA receptor modulator, a neurotransmitter reuptake inhibitor, a local anesthetic, an anticholinergic, a sodium channel blocker, a calcium channel blocker, a thyrotropin-releasing hormone, a y-secretase inhibitor, an AMPA receptor agonist or antagonist, an NMDA receptor agonist or antagonist, an mGlu receptor agonist or antagonist, a growth factor, an antiemetic agent, a corticosteroid; a cytotoxic agent; an antioxidant, an iron chelator, a mitochondrial modulator, a sirtuin modulator, a nitric oxide (NO) and/or nitric oxide synthase (NOS) modulator, a potassium channel agonist or antagonist, a purigenic receptor agonist or antagonist, and/or combinations thereof.

[0096] In some embodiments, the drug is selected from the group consisting of: benzocaine; carticaine; cinchocaine; cyclomethycaine; lidocaine; prilocaine; propxycaine; proparacaine;

tetracaine; tocainide; and trimecaine; methotrexate; cyclophosphamide; thalidomide; paclitaxel;

pemetrexed; pentostatin; pipobroman; pixantrone; plicamycin; procarbazine; raltitrexed;

rebeccamycin; rubitecan; SN-38; salinosporamide A; satraplatin; streptozotocin; swainsonine;

tariquidar; taxane; tegafur-uracil; temozolomide; testolactone; thioTEPA; tioguanine; topotecan;

trabectedin; tretinoin; triplatin tetranitrate; tris(2-chloroethyl)amine; troxacitabine; uracil mustard;

valrubicin; vinblastine; vincristine; vinorelbine; vorinostat; zosuquidar; carbamazepine;

oxcarbazepine; phenytein; valproic acid; sodium valproate; cinnarizine; flunarizine; nimodipine;

brain-derived neurotrophic factor (BDNF); ciliary neurotrophic factor (CNTF); glial cell-line derived

neurotrophic factor (GDNF); neurotrophin-3; neurotrophin-4; fibroblast growth factor (FGF)

receptor; insulin-like growth factor (IGF); and/or combinations thereof.

[0097] In some embodiments, provided is a method of delivering a photosensitizing agent to a human neuron or nerve comprising contacting the human neuron or nerve with a human neuron or

nerve targeting molecule comprising (a) a peptide that specifically binds to the neuron or nerve, or

component of either, and (b) a photosensitizing agent, wherein said targeting molecule comprises a

WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3),

DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20);

QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS (HNP401-N-8; SEQ ID

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124),

[0098] 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104), and/or combinations thereof.

[0099] In some embodiments, the method further comprises exposing the human neuron or nerve to a light source that activates the photosensitizing agent.

[00100] In some embodiments, the photosensitizing agent is selected from the group consisting of: a porphyrin, chlorin, and dye.

[00101] In some embodiments, the photosensitizing agent selected from the group consisting of: porphyrin, protoporfin IX, purlytin, verteporfin, HPPH, temoporfin, methylene blue,

photofrin, protofrin, hematoporphyrin, Talaporfin, benzopophyrin derivative monoacid, 5

aminileuvolinic acid, Lutetium texaphyrin, metallophthalocyanine, metallo

naphthocyaninesulfobenzo-porphyrazine, metallo-naphthalocyanines\, zinc tetrasulfophthalocyanine, bacteriochlorins, metallochlorins, chlorine derivative, Tetra(m

hydroxyphenyl)chlorin (mTHPC), pheophorbide, dibromofluorescein (DBF), IR700DX,

naphthalocyanine, porphyrin derivative, and/or combinations thereof.

[00102] In some embodiments, the human neuron or nerve targeting molecule is administered by systemic intravenous injection a human subject.

[00103] In some embodiments, the human neuron or nerve targeting molecule is administered prior to a surgical procedure. In some embodiments, the surgical procedure is a cancer surgical procedure. In some embodiments, the surgical procedure is a prostate cancer surgical procedure.

[00104] In some embodiments, provided is a pharmaceutical composition comprising: (a) a peptide that specifically binds to a human neuron, human nerve, or component of either, and (b) a

pharmaceutically acceptable excipient, wherein said human neuron or nerve targeting molecule

comprises a peptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID

NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3),

Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC

(HNP403 with GGC linker; SEQ ID NO:6), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac

QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC

(HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker;

SEQ ID NO:9), Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac

SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-SGQVPWEEPYYVVGGC

(HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker;

SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),

QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG

linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6;

SEQ ID NO:23); PYYVVKKSS (HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID

NO:25), SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQ ID

NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119), PYYVVKKSSGG

(HNP401-N-8 with GG linker; SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker;

SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),

SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG (HNP401-C-8 with

GG linker; SEQ ID NO:124), 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID

NO:104), and/or combinations thereof.

[00105] In some embodiments of the composition, the peptide is selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ

ID NO:2), and DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

[00106] In some embodiments of the composition, the peptide is selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401

ID NO:7), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20) QVPWEEPYYVVKKSSGG (HNP401-N-2

with GG linker; SEQ ID NO:21), and SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25).

[00107] In some embodiments of the composition, the peptide comprises SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1).

[00108] In some embodiments of the composition, the peptide comprises WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2).

[00109] In some embodiments of the composition, the peptide comprises DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

[00110] In some embodiments of the composition, the peptide comprises Ac SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4).

[00111] In some embodiments of the composition, the peptide comprises Ac WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5).

[00112] In some embodiments of the composition, the peptide comprises Ac DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6).

[00113] In some embodiments of the composition, the peptide comprises Ac QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7).

[00114] In some embodiments of the composition, the peptide comprises Ac PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8).

[00115] In some embodiments of the composition, the peptide comprises Ac EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9).

[00116] In some embodiments of the composition, the peptide comprises Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10).

[00117] In some embodiments of the composition, the peptide comprises Ac SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11).

[00118] In some embodiments of the composition, the peptide comprises Ac SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12).

[00119] In some embodiments of the composition, the peptide comprises Ac SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13).

[00120] In some embodiments of the composition, the peptide comprises Ac SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14).

[00121] In some embodiments of the composition, the peptide comprises DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16).

[00122] In some embodiments of the composition, the peptide comprises QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20).

[00123] In some embodiments of the composition, the peptide comprises QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).

[00124] In some embodiments of the composition, the peptide comprises PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22).

[00125] In some embodiments of the composition, the peptide comprises EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23).

[00126] In some embodiments of the composition, the peptide comprises PYYVVKKSS (HNP401-N-8; SEQ ID NO:24).

[00127] In some embodiments of the composition, the peptide comprises SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25).

[00128] In some embodiments of the composition, the peptide comprises SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26).

[00129] In some embodiments of the composition, the peptide comprises SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27).

[00130] In some embodiments of the composition, the peptide comprises SGQVPWEEP (HNP401-C-8; SEQ ID NO:28).

[00131] In some embodiments of the composition, the peptide comprises PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118).

[00132] In some embodiments of the composition, the peptide comprises EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119).

[00133] In some embodiments of the composition, the peptide comprises PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120).

[00134] In some embodiments of the composition, the peptide comprises SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121).

[00135] In some embodiments of the composition, the peptide comprises SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122).

[00136] In some embodiments of the composition, the peptide comprises SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123).

[00137] In some embodiments of the composition, the peptide comprises SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124).

[00138] In some embodiments of the composition, the peptide comprises 5FAM QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

[00139] In some embodiments of the composition, the peptide is SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1).

[00140] In some embodiments of the composition, the peptide is WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2).

[00141] In some embodiments of the composition, the peptide is DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

[00142] In some embodiments of the composition, the peptide is Ac SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4).

[00143] In some embodiments of the composition, the peptide is Ac WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5).

[00144] In some embodiments of the composition, the peptide is Ac-DLPDWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6).

[00145] In some embodiments of the composition, the peptide is Ac QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7).

[00146] In some embodiments of the composition, the peptide is Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8).

[00147] In some embodiments of the composition, the peptide is Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9).

[00148] In some embodiments of the composition, the peptide is Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10).

[00149] In some embodiments of the composition, the peptide is Ac SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11).

[00150] In some embodiments of the composition, the peptide is Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12).

[00151] In some embodiments of the composition, the peptide is Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13).

[00152] In some embodiments of the composition, the peptide is Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14).

[00153] In some embodiments of the composition, the peptide is DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16).

[00154] In some embodiments of the composition, the peptide is QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20).

[00155] In some embodiments of the composition, the peptide is QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).

[00156] In some embodiments of the composition, the peptide is PWEEPYYVVKKSS (HNP401 N-4; SEQ ID NO:22).

[00157] In some embodiments of the composition, the peptide is EEPYYVVKKSS (HNP401-N 6; SEQ ID NO:23).

[00158] In some embodiments of the composition, the peptide is PYYVVKKSS (HNP401-N-8; SEQ ID NO:24).

[00159] In some embodiments of the composition, the peptide is SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25).

[00160] In some embodiments of the composition, the peptide is SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26).

[00161] In some embodiments of the composition, the peptide is SGQVPWEEPYY (HNP401-C 6; SEQ ID NO:27).

[00162] In some embodiments of the composition, the peptide is SGQVPWEEP (HNP401-C-8; SEQ ID NO:28).

[00163] In some embodiments of the composition, the peptide is 5FAM QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

[00164] In some embodiments of the composition, the peptide is PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118).

[00165] In some embodiments of the composition, the peptide is EEPYYVVKKSSGG (HNP401 N-6 with GG linker; SEQ ID NO:119).

[00166] In some embodiments of the composition, the peptide is PYYVVKKSSGG (HNP401-N 8 with GG linker; SEQ ID NO:120).

[00167] In some embodiments of the composition, the peptide is SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121).

[00168] In some embodiments of the composition, the peptide is SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122).

[00169] In some embodiments of the composition, the peptide is SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123).

[00170] In some embodiments of the composition, the peptide is SGQVPWEEPGG (HNP401 C-8 with GG linker; SEQ ID NO:124).

[00171] In some embodiments of the composition, the peptide is bound to a cargo. In some embodiments, the cargo is a drug, photosensitizing agent, or fluorescent moiety.

BRIEF DESCRIPTION OF THE DRAWINGS

[00172] Figure 1: Fluorescence images of exposed sciatic nerves in living wild type mice following administration of 450nmols HNP401 in three mice (left) and NP41 (two mice, right). Both

left and right sciatic nerves are shown. FAM is the fluorescein attached to the C-terminal lysine of

each peptide sequence. Images were obtained with a Zeiss Lumar.

[00173] Figure 2: Quantitation of images from figure 1 show HNP 401 have similar contrast (nerve fluorescent intensity/muscle fluorescent intensity) to NP41 (6.2 fold compared to 6.7 fold)

with with higher intensity labeling of both nerve and adjacent muscle tissue. 6 nerves, 3 mice total

for HNP401 and 2 mice (4 nerves) for NP41.Y axis = average fluorescent (515nm emission) intensity

calculated in imageJ.

[00174] Figure 3: Topical application of Nerve binding peptides on sections of human nerve showing high binding of HNP401. Exposure gain was decrease (30 to 10) for HNP401 as exposure

under identical settings, compared to other standards, were saturated by high signal.

[00175] Figure 4: Fluorescence from sectioned human nerve after topical application of NP41, HNP401 and HNP404. Peptides were applied at 100 uM for 20 mins followed by washing in

PBS and imaging on a Nikon Al confocal microscope. All images are leveled equally.

[00176] Figure 5: In-vivo labeling of Rat Sciatic nerve with HNP401

[00177] Figure 6: Fluorescent labeling of rat prostate cavernosal nerve in live rats. HNP301 is an early generation nerve binding peptides that is not showing as much contrast for prostate nerve

labeling compared to HNP401.

[00178] Figure 7: In-vivo labeling of prostatic neurovascular bundle with HNP401. HNP401 labeling of autonomic nerve bundles in live rats.

[00179] Figure 8: Screening of HUMAN nerve binding peptides identified by phage display. Topical application of 100mM of human nerve binding peptides FAM-HNP401 (A), FAM-HNP402 (B),

FAM-HNP403 (C) on serial sections of fresh-viable human sural nerve (top image) and human temporalis muscle (bottom image). For comparison topical application of 100mM of carboxy-FAM

(D) and peptide screened for binding to mouse nerve NP41-FAM (E). H&E of staining of nerve and

muscle (F). All fluorescence images acquired on Lumar microscope at 34X magnification with a 2s

exposure and levelled equally for comparison. NTQTLAKAPEHT (NP41; SEQ ID NO:15 from U.S.

Patent No. 8,685,372 or International Patent Publication No. W2010121023A2).

[00180] Figure 9: Comparison of FAM-HNP401 and FAM-NP41 in binding and labelling of HUMAN sural nerve. Topical application of 100mM of HNP401-FAM on 10mm sections of unfixed

human brachial plexus nerve tissue (A) and human temporalis muscle tissue (D) kept adjacent on

same glass slide and imaged on a confocal microscope with 488nm excitation laser. For comparison,

NP41-FAM was applied to human nerve (B) and muscle (C) under identical conditions as mentioned

for (A and D). H&E staining of the nerve (C) and muscle (F). Signal intensity of perineurium of nerve

tissue treated with HNP401-FAM (n=4) compared with NP41-FAM (n=4) (G). Nerve to muscle

contrast of peptides applied topically to human tissue sections (n=4) (H).

[00181] Figure 10: Differential binding of nerve binding peptides to HUMAN and MOUSE tissue. Human tissue: Determination of optimal dose response by topical application of HNP401

FAM on human sural nerve sections at final concentration of 375 mM (A), 100 mM (B), 50 mM (C),

10 mM (D) and 1 mM (E), imaged with confocal microscopy with identical parameters and levelled

equally for comparison. ** brightened 2 fold for viewing. Nerve and muscle contrast at high

concentration of 375 mM for FAM-NP41 (F and G) and FAM-HNP401 (H and 1) imaged on confocal

microscopy and levelled for direct comparison. Mouse tissue: Mouse facial nerve (red arrows) with

surrounding muscle treated with 375 mM (J), 100 mM (K) of FAM-NP41 or 375 mM (L), 100 mM (M)

of FAM-HNP401. Images in bottom row acquired on Lumar imaging scope with identical parameters

and are comparable. FAM-HNP401 shows high binding of muscle in mouse tissue with poor contrast

compared to FAM-NP41.

[00182] Figure 11: In-vivo imaging of nerve binding peptides in RODENTS with pharmacokinetic profile following IV injection. In-vivo fluorescence image of sciatic nerve of 6

month old SKH1-Elite mice injected i.v. with 450nmols of FAM-HNP401 (A) or FAM-NP41 (B) and

imaged on the Lumar imaging scope 2h post injection. Intensity of sciatic nerve was measured in

Image J shows a 2.3 fold increase in binding for peptide screened for binding human nerve (HNP401)

vs peptide screened for binding to mouse nerve (NP41) (C). However, the nerve to surrounding

muscle contrast for the two peptides are comparable at 5.79±0.81for FAM-HNP401 and 6.63 ±1.63

for FAM-NP41 in mouse thigh (D). In-vivo fluorescence image of prostate nerve plexus using real

time custom surgical imaging system (E) and Lumar small animal microscope (F) 5 hours after t.v.

injection of 2pmoles of HNP401-FAM in 100gm male Sprague Dawley rat. Sciatic nerve in rat was imaged 5 hours after systemic injection of 2pmoles of FAM-HNP401 (G). Blood clearance curve

shows FAM signal obtained from equal volume of blood draws taken from five SKH1-Elite male mice.

Each mouse was injected i.v. with100nmol of FAM-HNP401 prior to blood collection at1min, 10min,

20min, 30min, 1h and 2h timepoints (H).

[00183] Figure 12: HNP401 binds to HUMAN nerves (cavernosal and median ante-brachial cutaneous) Fluorescent imaging after topical application of 100pM FAM-HNP401 or FAM-NP41 on

10pm sections on cryosectioning tape of nerve within human prostate gland, (top row, A and B) or

from median anti-brachial cutaneous human nerve (bottom row, B and F). Nerves were imaged

immediately after sectioning and application of peptide using confocal microscopy.

Immunohistochemistry analysis with dual label for neurofilament antibody SM1312 (red) and DAPI stained nuclei (blue) (C and G) of fixed section of nerve and corresponding H&E staining (D and H) on glass slides.

[00184] Figure 13: HNP401 binds to HUMAN cavernosal nerves. Topical application of 100pM FAM-HNP401 (A) or FAM-NP41(B) on 10lm sections on cryosectioning tape of unfixed fresh

viable nerves from the prostate gland, using confocal microscopy. Immunofluorescence (C)

neurofilament antibody SM1312 on fixed section of nerve from prostate gland and corresponding

H&E staining (D) on glass slides. These images are obtained from different patients than those

shown in Fig 5. of this document.

[00185] Figure 14: Determination of HUMAN nerve binding domain of HNP401 by sequential deletion. Representative images fluorescence images of unfixed human sural nerve

treated topically with 1OOuM of FAM labelled HNP401 (A), HNP401-N-2 (B), HNP401-N4 (C),

HNP401-N6 (D), HNP401-N8 (E), HNP401-N4 C-2 (F), HNP401-N4 C-4 (G), HNP401-N4 C-6 (H),

HNP401-N4 C-8 (1).

[00186] Figure 15: Quantitation of nerve binding of deletion variants of HNP401. Quantitation of nerve binding of each HNP401 variant shown in figure 7 (n=5).

[00187] Figure 16: In-vivo fluorescent labelling of autonomic nerve in rodent. Low magnification fluorescent image showing bladder, vas deferens and urethra running through the

prostate with adjacent autonomic nerve labeled with FAM-NP41 in mice (A). Higher magnification

white light reflectance image (B) and corresponding fluorescence grayscale image (C) of the

autonomic nerve running adjacent to the urethra. Quantitation of autonomic nerve detection by

fluorescence compared to white light detection in mice (D) Nerve to muscle contrast for

reflectance/fluorescence were plotted for individual nerve branches. Values to the right of the line indicate that there is improved visualization with fluorescence compared to reflected light. Images

(E-G) are analogous to (A-C) except they highlight FAM-NP41 dependent labeling of autonomic nerve

in rat prostate versus mouse, with white light imaging showing non-visible nerve (F). FAM-NP41

labeled prostate nerve is also detectable using a clinical grade Zeiss Pentero Surgical Microscope (H).

[00188] Figure 17: Comparison of FAM-HNP401 and FAM-NP41 in binding and labelling of human sural nerve. Topical application of 100 M of FAM-HNP401 on 10lm sections of unfixed

human sural nerve tissue (A) and human temporalis muscle tissue (E) kept adjacent on same glass

slide and imaged on a confocal microscope with 488nm excitation laser. For comparison, FAM-NP41 was applied to a sequential section of human nerve (B)and muscle (F) under identical conditions as mentioned for (A and E). H&E staining of the nerve (C) and muscle (G). Signal intensity of perineurium of nerve tissue treated with FAM-HNP401 (n=4) compared with FAM-NP41 (n=4) (D). Nerve to muscle contrast of peptides applied topically to human tissue sections (n=4) (H).

[00189] Figure 18: In-vivo imaging of nerve binding peptides in mouse and rat with pharmacokinetics. In-vivo fluorescence image of sciatic nerves from 6 month old SKH1 mice that had

been previously intravenously injected with 450nmols of FAM-HNP401 [~48.4 mg/kg] (A) or FAM

NP41 [~39mg/kg] (B). Intensity of sciatic nerve measured and quantitated in Image J showed a 2.3 fold increase for FAM-HNP401 compared to FAM-NP41(C). Nerve to muscle contrast for the two

peptides were comparable at 5.79±0.81 for FAM-HNP401 and 6.63±1.63 for FAM-NP41 (D). In-vivo

fluorescence image of rat sciatic nerve 5 hours after intravenous injection of 2pmoles of FAM

HNP401 [~54mg/kg] (E). Rat prostate nerve imaged with real time custom surgical imaging system (F)

and Lumar small animal microscope (G) 5 hours after intravenous injection of 2pmoles of FAM

HNP401. Blood clearance curve shows FAM signal obtained from equal volume of blood draws taken

from five SKH1-Elite male mice (H). Each mouse was injected intravenous with 100nmol [~11mg/kg]

of FAM-HNP401 prior to blood collection at 1min, 10min, 20min, 30min, 1h and 2h time points.

[00190] Figure 19: HNP401 binds to fresh viable nerve from prostate gland and median anti-brachial cutaneous human nerve. Fluorescent imaging after topical application of 100pM FAM

HNP401 or FAM-NP41 on 10pm sections on cryosectioning tape of nerve within human prostate

gland, (top row, A and B) or from median anti-brachial cutaneous human nerve (bottom row, E and

F). Nerves were imaged immediately after sectioning and application of peptide using confocal

microscopy. Immunohistochemistry analysis with dual label for neurofilament antibody SM1312 (red)

and DAPI stained nuclei (blue) (C and G) of fixed section of nerve and corresponding H&E staining (D

and H) on glass slides.

[00191] Figure 20: Comparison of truncated sequences to determine binding efficiency. Representative images fluorescence images of unfixed human sural nerve that were treated topically

with 100pM of FAM labelled N-2 (A), N-4 (B), N-6 (C), N-8 (D), C-2 (E), C-4 (F), C-6 (G), C-8 (H) or

HNP401 (1). Due to poor solubility C-6 had a final concentration of *731M and C-8 had a final

concentration of **80.6pM for topical tests. Comparison of signal intensity of peptides normalized to

FAM-HNP401 were made to test for improved binding (J). Normalized sural nerve to temporalis muscle contrast was determined for FAM-HNP401 and FAM-HNP401-N-2 (Student t-test, unpaired, one-tail, p=0.011) (K).

[00192] Figure 21: Food dyes efficiently quench FAM-NP41 bladder fluorescence. A fluorescent plate reader assay was used to show dose dependent quenching of FAM-NP41

fluorescence. Erythrosine extra bluish (Santa Cruz Biotechnology, Inc.) was the most efficient

quencher with > ~80% quenching at 2.5 times dye to fluorescein ratio and > 95% quenching at 5X

dye to fluorescein ratio (A). Other food dyes tested included Allura Red and Sunset Yellow. To test

for quenching in-vivo we administered, by direct iv injection, 50 mg/kg MW 879.76 (~1.5 moles per 25 gm mouse) to mice that had been injected with 150nmoles of FAM-NP41 2 hours prior. This

represents approximately a 10 X dye to FAM-NP41 dose. Some bladder fluorescence remained after

imaging so additional dye (30 Ipl, 10mM Erythrosin extra bluish) was injected directly into the

bladder. Images are shown for mouse bladder with no dye quencher (B) and addition of Erythrosine

extra bluish (intravenous and intra bladder) (C) with bladder fluorescence quench to near

background level. Dye would likely not be needed if this method was used for human patients as

bladder catheterization in patients could be started as FAM-NP41 is administered so bladder

fluorescence could be washed out.

[00193] Figure 22: TAMRA-NP41labels autonomic unmyelinated nerve in prostate gland of rat. Fluorescence image of nerve fascicles (white arrows) around prostate gland in living male

Sprague-Dawley rat, imaged 15 min post i.v. injection of 500nmol of NP41-TAMRA (A). Tissue was

excised and frozen unfixed for validation of peptide fluorescence signal using confocal imaging (B)

and immunohistochemistry with an antibody to TAMRA detected with horse radish peroxidase

secondary and diaminobenzidine staining (C). Antibody staining against tyrosine hydroxylase was

used tovalidate presence of autonomic nerves (D) no-primary negative control (E).

[00194] Figure 23: Screening of human nerve binding peptides identified by phage display. Topical application of 100ptM of human nerve binding peptides FAM-HNP401 (A), FAM-HNP402 (B),

FAM-HNP403 (C) on serial sections of fresh-viable human sural nerve (upper row) and human

temporalis muscle (lower row). For comparison topical application of 100M of carboxy-FAM (D) and

peptide screened for binding to mouse nerve NP41-FAM (E). H&E of staining of nerve and muscle (F).

All fluorescence images acquired on Lumar microscope at 34X magnification with a 2s exposure and

levelled equally for comparison.

[00195] Figure 24: Screening of human nerve binding peptides identified by phage display. Topical application of 100ptM of human nerve binding peptides FAM-HNP401 (A), FAM-HNP402 (B),

FAM-HNP403 (C) on serial sections of fresh-viable human ansa cervicalis nerve (upper row) and

human great auricular nerve (lower row) from the neck of two different patients. For comparison

topical application of 100pM of carboxy-FAM (D) and peptide screened for binding to mouse nerve

NP41-FAM (E). All fluorescence images acquired on Lumar microscope at 34X magnification with a 2s

exposure and levelled equally for comparison.

[00196] Figure 25: Differential binding of nerve binding peptides to human and mouse tissue. Determination of optimal dose response by topical application of FAM-HNP401 on human

laryngeal nerve sections at final concentration of 375 pM (A), 100 pM (B), 50 pM (C), 10 pM (D) and

1 M (E), imaged with confocal microscopy with identical parameters and levelled equally for

comparison. ** brightened 2 fold for viewing. Nerve to muscle contrast at high concentration of 375

aM for FAM-NP41 (F and G) and FAM-HNP401 (H and 1) imaged on confocal microscopy and levelled

for direct comparison. Mouse facial nerve (red arrows) with surrounding muscle treated with 375

IiM (J), 100 IiM (K) of FAM-NP41 or 375 IiM (L), 100 IiM (M) of FAMHNP401. Images in bottom row acquired on Lumar imaging scope with identical parameters and are comparable. FAM-HNP401

shows high binding of muscle in mouse tissue with poor contrast compared to FAMNP41. High

resolution confocal image of low concentration of FAM-HNP401 (10 pM) on human nerve shows

binding of peptide to non-axonal structural components of nerve (N).

[00197] Figure 26: Autofluorescence of human nerve tissue. Topical application of 100.IM FAM-HNP401 (A) or buffer only (B) on 10lm sections of unfixed human sural nerve followed by

imaging using confocal microscopy under identical acquisition parameters for direct comparison.

Images were levelled equally using Image J followed by a 16 fold brightening of (B) for viewing.

[00198] Figure 27: Mass spectroscopy analysis of urine samples from mice injected with nerve binding peptides. Fragmented ion peaks from Cysteine-FAM collected from the urine of mice

that were injected with FAM-HNP401 indicating peptide is metabolized (A). Similar results were

obtained with mice injected with FAM-NP41 (B). However, mouse injected with FAM-dNP41, where

peptide is made with d-amino acids, is detectable in the urine and is not metabolized (C).

[00199] Figure 28: Stability of peptides in ex-vivo human plasma and cerebrospinal fluid from rats. FAM-HNP401 peptide detected at 5min (A) and 2hours (B) after incubation at 37o C in

human plasma in at a dose of 53.2 mg/kg or 2mole. An equal volume of 1:1 acetonitirile: water with 2% acetic acid is added to precipitate the protein matter, supernatant is extracted for analysis by LC-MS on a C18 reverse phase column with gradient of 9:1 H2+0.05%TFA:

Acetonitrile+0.05%TFA to 1:9 H20+0.05%TFA: Acetonitrile+0.05%TFA in 20min. Detector channel of 450nm shows FAM-HNP401. The peptide remain intact at 2hours post incubation with x% of the

composition at 5min post incubation with human plasma. FAM-NP41 peptide detected at 5min (C)

and 2hours (D) after incubation at 37o C in human plasma in at a dose of 53.2 mg/kg or 2pmole,

followed by LC-MS analysis with method described above. Similar to our previous result, FAM-NP41

remains intact at 2hours post incubation with x% of the composition at 5min post incubation with

human plasma. FAM-HNP401 (E) and FAM-NP41 (F) were also tested in cerebrospinal spinal fluid

from rat at 2 hours after incubation to demonstrate stability of the peptides in circulation.

[00200] Figure 29: FAM-HNP401 binds to fresh viable nerve acquired from human prostate gland. Topical application of 100ptM FAM-HNP401 (A) or FAM-NP41 (B) on 10lm sections of unfixed

nerves from the prostate gland followed by imaging using confocal microscopy.

Immunofluorescence for nerve using neurofilament antibody SM1312 (C) on fixed section of nerve

from prostate gland and corresponding H&E staining (D). These images are obtained from different

patients than those shown in Figure 20.

[00201] Figure 30: Table of peptide sequences and their abbreviations.

DETAILED DESCRIPTION OF THE INVENTION

I. Introduction

[00202] Disclosed herein, in certain embodiments, are targeting molecules comprising a peptide that specifically binds to a human neuron, human nerve, or component of either. In some

embodiments, the peptide is selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP

401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403;

SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC

(HNP401 with GGC linker; SEQ IDNO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker;

SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac

QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ IDNO:7), Ac-PWEEPYYVVKKSSGGC

(HNP401-N-4 with GGC linker; SEQ IDNO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker;

SEQ ID NO:9), Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac

SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG

linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6;

SEQ ID NO:23), PYYVVKKSS (HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID

NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker;

SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119), PYYVVKKSSGG

SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),

GG linker; SEQ ID NO:124), and 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ

ID NO:104). In some embodiments, the peptide is not Ac-SHSNTQTLAKAPEHTGC (Ac-NP41; SEQ ID

NO:17). In some embodiments, the peptide is not SHSNTQTLAKAPEHTGC (NP41; SEQ ID NO:18). In

some embodiments, the peptide is not NTQTLAKAPEHT (NP41; SEQ ID NO:19).

II. Definitions

[00203] In the present description, any concentration range, percentage range, ratio range, or integer range is to be understood to include the value of any integer within the recited range and,

when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless

otherwise indicated. Also, any number range recited herein relating to any physical feature, such as

polymer subunits, size or thickness, are to be understood to include any integer within the recited

range, unless otherwise indicated. As used herein, the term "about" means ±20% of the indicated

range, value, or structure, unless otherwise indicated.

[00204] It should be understood that the terms "a" and "an" as used herein refer to "one or more" of the enumerated components. The use of the alternative (e.g., "or") should be understood

to mean either one, both, or any combination thereof of the alternatives.

[00205] In addition, it should be understood that the individual compounds, or groups of compounds, derived from the various combinations of the structures and substituents described

herein, are disclosed by the present application to the same extent as if each compound or group of compounds was set forth individually. Thus, selection of particular structures or particular substituents is within the scope of the present disclosure

[00206] As used herein, the following terms have the meanings ascribed to them unless specified otherwise.

[00207] The central nervous system (CNS) consists of the brain and the spinal cord, as well as the retina.

[00208] The peripheral nervous system (PNS) extends outside the CNS. The PNS is divided into the somatic nervous system and the autonomic nervous system.

[00209] A neuron is an electrically excitable cell that processes and transmits information by electrical and chemical signaling.

[00210] A typical neuron possesses a cell body (often called the soma), dendrites, and an axon.

[00211] A nerve is an enclosed, cable-like bundle of neural axons. Each nerve is a cordlike structure that contains many axons. Each axon is surrounded by a layer of tissue called the

endoneurium. The axons are bundled together into groups called fascicles, and each fascicle is

wrapped in a layer of tissue called the perineurium. The neuron or nerve is wrapped in a layer of

tissue called the epineurium.

[00212] As used herein, the term "targeting molecule" refers to any agent (e.g., peptide, protein, nucleic acid polymer, aptamer, or small molecule) that specifically binds to a target of

interest. In some embodiments, the targeting molecule comprises a peptide, also referred to herein

as "targeting peptide." The target of interest may be a tissue, a cell type, a cellular structure (e.g., an

organelle), a protein, a peptide, a polysaccharide, or a nucleic acid polymer. In some embodiments,

the targeting molecule is any agent that specifically binds to one or more neurons or nerves of a

subject. In some embodiments, the targeting molecule further comprises a cargo (e.g., drug, fluorescent label, or photosensitizing agent).

[00213] As used herein, the term "aptamer" refers to an oligonucleotide (e.g., DNA, RNA, or XNA) molecule that has been selected from random pools based on their ability to bind other

molecules with high affinity specificity based on non- Watson and Crick interactions with the target molecule (see, e.g., Cox and Ellington, Bioorg. Med. Chem. 9:2525-2531 (2001); Lee et al, Nuc. Acids

Res. 32:D95-D100(2004)) or a short peptide (e.g., 5-20 amino acids) that is embedded as a loop

within a stable protein scaffold rather than as a free peptide. Aptamers can be selected which bind nucleic acid, proteins, small organic compounds, vitamins, inorganic compounds, cells, and even

entire organisms. In some embodiments, the targeting peptide can comprise an aptamer or the

targeting molecule peptide sequence can be in the format of an peptide aptamer.

[00214] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to naturally occurring occurring amino acid polymers as well as amino acid polymers in which one or more amino acid residues is a non

naturally occurring amino acid (e.g., an amino acid analog). The terms encompass amino acid chains

of any length, including full length proteins (i.e., antigens), wherein the amino acid residues are

linked by covalent peptide bonds. As used herein, the term "peptide" refers to a polymer of amino

acid residues typically ranging in length from 2 to about 50 residues. In certain embodiments the

peptide ranges in length from about 2, 3, 4, 5, 7, 9, 10, or 11 residues to about 50, 45, 40, 45, 30, 25,

20, or 15 residues. In certain embodiments the peptide ranges in length from about 8, 9, 10, 11, or

12 residues to about 15, 20 or 25 residues. Where an amino acid sequence is provided herein, L-, D-,

or beta amino acid versions of the sequence are also contemplated as well as retro, inversion, and

retro-inversion isoforms. Peptides also include amino acid polymers in which one or more amino

acid residues is an artificial chemical analogue of a corresponding naturally occurring amino acid, as

well as to naturally occurring amino acid polymers. In addition, the term applies to amino acids

joined by a peptide linkage or by other modified linkages (e.g., where the peptide bond is replaced by an a-ester, a /3-ester, a thioamide, phosphonamide, carbamate, hydroxylate, and the like (see,

e.g., Spatola, (1983) Chem. Biochem. Amino Acids and Proteins 7: 267-357), where the amide is

replaced with a saturated amine (see, e.g., Skiles et al., U.S. Pat. No. 4,496,542, which is

incorporated herein by reference, and Kaltenbronn et al., (1990) Pp. 969-970 in Proc. 1Ith American

Peptide Symposium, ESCOM Science Publishers, The Netherlands, and the like)).

[00215] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the

naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic

code, as well as those amino acids that are later modified, e.g., hydroxyproline, y-carboxyglutamate,

and O-phosphoserine. An amino acid may be an L- or D-amino acid. Amino acid analogs refer to

compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., a carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics refers to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally occurring amino acid.

[00216] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUP AC-UB Biochemical Nomenclature Commission. Nucleotides, likewise, may be referred to by their commonly accepted single-letter

codes.

[00217] One of skill will recognize that individual substitutions, deletions or additions to a peptide, polypeptide, or protein sequence which alters, adds or deletes a single amino acid or a

small percentage of amino acids in the encoded sequence is a "conservatively modified variant"

where the alteration results in the substitution of an amino acid with a chemically similar amino

acid. Conservative substitution tables providing functionally similar amino acids are well known in

the art. Such conservatively modified variants are in addition to and do not exclude polymorphic

variants, interspecies homologs, and alleles of the invention.

[00218] The following eight groups each contain amino acids that are conservative substitutions for one another: 1) Alanine (A), Glycine (G); 2) Aspartic acid (D), Glutamic acid (E); 3)

Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (1), Leucine (L), Methionine

(M), Valine (V); 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W); 7) Serine (S), Threonine (T); and 8)

Cysteine (C), Methionine (M) (see, e.g., Creighton, Proteins (1984)).

[00219] "Sequence identity," as used herein, refers to the percentage of amino acid residues in a single sequence that are identical with the amino acid residues in another reference polypeptide

sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum

percent sequence identity, and not considering any conservative substitutions as part of the

sequence identity. The percentage sequence identity values can be generated using the NCBI BLAST

2.0 software as defined by Altschul et al. (1997), Nucl. Acids Res. 25:3389-3402, with the parameters

set to default values.

[00220] As used herein, the terms "label" refers to a molecule that facilitates the visualization and/or detection of a targeting molecule disclosed herein. In some embodiments, the

label is a fluorescent moiety.

[00221] The phrase "specifically binds" when referring to the interaction between a targeting molecule disclosed herein and a target (e.g., purified protein, neuron or nerve tissue, neuron or

nerves, cranial neuron or nerves, central neuron or nerves, myelinated or unmyelinated neuron or

nerves, or connective tissue surrounding neuron or nerves), refers to the formation of a high affinity

bond between the targeting molecule and the target. Further, the term means that the targeting molecule has low affinity for non-targets.

[00222] "Selective binding," "selectivity," and the like refer to the preference of agent to interact with one molecule as compared to another. Preferably, interactions between a targeting

molecule disclosed herein and a target are both specific and selective. Note that in some

embodiments an agent is designed to "specifically bind" and "selectively bind" two distinct, yet similar targets without binding to other undesirable targets.

[00223] The terms "individual," "patient," or "subject" are used interchangeably. As used herein, they mean any mammal (i.e. species of any orders, families, and genus within the taxonomic

classification animalia: chordata: vertebrata: mammalia). In some embodiments, the mammal is a

cow, horse, sheep, pig, cat, dog, goat, mouse, rat, rabbit, guinea pig, non-human primate, or human. None of the terms require or are limited to situation characterized by the supervision (e.g. constant

or intermittent) of a health care worker (e.g. a doctor, a registered nurse, a nurse practitioner, a

physician's assistant, an orderly, or a hospice worker).

[00224] The terms "administer," "administering", "administration," and the like, as used herein, refer to the methods that may be used to enable delivery of agents or compositions to the desired site of biological action. These methods include, but are not limited to parenteral injection

(e.g., intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular, intrathecal,

intravitreal, infusion, or local). Administration techniques that are optionally employed with the

agents and methods described herein, include e.g., as discussed in Goodman and Gilman, The

Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical

Sciences (current edition), Mack Publishing Co., Easton, Pa. In some embodiments, administration is

via systemic intravenous injection into human patients.

[00225] The term "pharmaceutically acceptable" as used herein, refers to a material that does not abrogate the biological activity or properties of the agents described herein, and is relatively nontoxic (i.e., the toxicity of the material significantly outweighs the benefit of the material). In some instances, a pharmaceutically acceptable material may be administered to an individual without causing significant undesirable biological effects or significantly interacting in a deleterious manner with any of the components of the composition in which it is contained.

[00226] The term "surgery" as used herein, refers to any methods for that may be used to manipulate, change, or cause an effect by a physical intervention. These methods include, but are not limited to open surgery, endoscopic surgery, laparoscopic surgery, minimally invasive surgery, robotic surgery, any procedures that may affect any neuron or nerves such as placement of retractors during spinal surgery, cardiac neuron or nerve ablation, epidural injection, intrathecal injections, neuron or nerve blocks, implantation of devices such as neuron or nerve stimulators and implantation of pumps. In some embodiments, the subject of the surgery is a human subject or human patient

Ilil. Targets

[00227] Disclosed herein, in certain embodiments, are human neuron and/or nerve targeting molecules that specifically bind to a human neuron or nerve target.

[00228] In some embodiments, the target is a human neuron or nerve. The nerve is any human nerve (e.g., motor nerves, sensory nerves, sympathetic and parasympathetic nerves, periprostatic neurovascular bundle, sciatic nerves, cranial nerves including olfactory nerve, optic nerve, oculomotor nerve, trochlear nerve, trigeminal nerve, abducens nerve, facial nerve, vestibulocochlear nerve, glossopharyngeal nerve, vagus nerve, accessory nerve, hypoglossal nerve, spinal nerves, brachial plexus, or lumbrosacral plexus). The neuron is any neuron (e.g., sensory neurons (afferent neurons), motor neurons (efferent neurons), interneurons, unipolar neurons, bipolar neurons, multipolar neurons, basket cells, Betz cells, medium spiny neurons, Purkinje cells, pyramidal cells, Renshaw cells, Granule cells, anterior horn cells). In some embodiments, the human neuron or nerve is myelinated. In some embodiments, the neuron or nerve is unmyelinated. In some embodiments, the human neuron or nerve isdemyelinated. In some embodiments, the human neuron or nerve is undergoing demyelination.

[00229] In some embodiments, the neuron and/or nerve target is a component of a human neuron or nerve. The component of a human neuron or nerve is any component of a neuron or nerve. In some embodiments, the target is tissue within or surrounding a neuron or nerve (e.g., epineurium, perineurium, or endoneurium). In some embodiments, the target is a component of myelin, (e.g., myelin basic protein (MBP), myelin oligodendrocyte glycoprotein, or proteolipid protein). In some embodiments, the target is expressed by Schwann cells, (e.g., MBP, glial fibrillary acidic protein, S-100, or myelin protein zero). In some embodiments, the target is a component of neuron or nerve tissue, (e.g., elastin, fibrillin, e-cadherin, cytokeratin, vimentin, collagenI, collagen, III, collagen IV, or collagen V). In some embodiments, the target is a neurotrophic factor receptor expressed in neuron or nerves, (e.g., tyrosine kinase receptors TrkA, TrkB, and TrkC, low affinity neuron or nerve growth receptor or p75 neurotrophin receptor, or GDNF family receptor alpha- 1 or -2). In some embodiments, the target is a non-neurotrophic factor receptor expressed in a neuron or nerve tissue, (e.g., epithelial growth factor receptors, transforming growth factor beta receptors, vascular endothelial growth factor receptors, endothelin A receptors, endothelin B receptors, and integrin receptors).

[00230] Determining whether a neuron and/or nerve targeting molecule is capable of binding a human neuron or nerve or component thereof is accomplished by any suitable method. In some embodiments, the method of determining whether a neuron and/or nerve targeting molecule is capable of binding a human neuron or nerve or component thereof involves contacting a targeting molecule (e.g., peptide or aptamer) disclosed herein with a test agent for a period of time sufficient to allow the targeting molecule and test agent to form a binding complex. The binding complex is detected using any suitable method. Suitable binding assays can be performed in vitro or in vivo and include, but are not limited to, phage display, two-hybrid screens, co-precipitation, cross-linking, and expression cloning (see, e.g., Bennet, J.P. and Yamamura, H.I. (1985) "Neurotransmitter, Hormone or Drug Receptor Binding Methods," in Neurotransmitter Receptor Binding (Yamamura, H. L, et al., eds.), pp. 61-89. Other binding assays involve the use of mass spectrometry or NMR techniques to identify molecules bound to the target of interest. The targeting molecule utilized in such assays can be naturally expressed, cloned or synthesized.

[00231] In some embodiments, the targeting molecule is capable of crossing the blood-brain barrier in order to reach and bind the human neuron or nerve of interest.

IV. Targeting Molecules Peptides and Aptamers

[00232] Provided in the present disclosure are peptides that bind to human motor/sensory and autonomic nerves and can be used in human neuron or nerve targeting molecules of the present

invention. In some embodiments, a targeting peptide comprises an amino acid sequence of

SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2),

DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac

SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC

(HNP402 with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac

PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac-EEPYYVVKKSSGGC (HNP401-N

6 with GGC linker; SEQ ID NO:9), Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10),

Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-SGQVPWEEPYYVVGGC

SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),

(HNP401-N-8 with GG linker; SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),

GG linker; SEQ ID NO:124), or 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ

ID NO:104).

[00233] In some embodiments, the human neuron or nerve targeting molecule comprises a peptide sequence selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID

DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC

linker; SEQ ID NO:11), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20);

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), and 5FAM

QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104). In some embodiments,

peptides of the present invention comprise an amino acid sequence of about 8 to about 25 amino acids (e.g., 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids), about 10

to about 23 amino acids, or about 15 to about 21 amino acids comprising the core binding domain of

PYYVVKK (SEQ ID NO:40). In some embodiments, peptides of the present invention comprise an

amino acid sequence of about 13 to about 25 amino acids comprising the core binding domain of

PYYVVKK (SEQ ID NO:40) and an N-terminal sequence of QVPWEE (SEQ ID NO:41). In some

embodiments, the peptides of the present invention comprise an amino acid core binding domain of

PYY (SEQ ID NO:116) or PYYVV (SEQ ID NO:117) and an N-terminal sequence of QVPWEE (SEQ ID

NO:41). In some embodiments, the peptides of the present invention comprise an amino acid core

binding domain of PYY (SEQ ID NO:116) and an N-terminal sequence of QVPWEE (SEQ ID NO:41). In

some embodiments, the peptides of the present invention comprise an amino acid core binding

domain of PYYVV (SEQ ID NO:117) and an N-terminal sequence of QVPWEE (SEQ ID NO:41).

[00234] One such embodiment is a peptide of QVPWEEPYYVVKK (SEQ ID NO:42). In some embodiments, the targeting molecule comprises a peptide that is not Ac-SHSNTQTLAKAPEHTGC (Ac

NP41 with GC linker; SEQ ID NO:17). In some embodiments, the targeting molecule comprises a

peptide that is not SHSNTQTLAKAPEHTGC (NP41 with GC linker; SEQ ID NO:18). In some

embodiments, the peptide is not NTQTLAKAPEHT (NP41; SEQ ID NO:19).

[00235] In some embodiments the targeting molecule comprises a peptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

[00236] In some embodiments, the targeting molecule comprises a peptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), and QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).

[00237] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1).

[00238] In some embodiments, the targeting molecule comprises the peptide WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2).

[00239] In some embodiments, the targeting molecule comprises the peptide DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3).

[00240] In some embodiments, the targeting molecule comprises the peptide Ac SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4).

[00241] In some embodiments, the targeting molecule comprises the peptide Ac WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5).

[00242] In some embodiments, the targeting molecule comprises the peptide Ac DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6).

[00243] In some embodiments, the targeting molecule comprises the peptide Ac QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7).

[00244] In some embodiments, the targeting molecule comprises the peptide Ac PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8).

[00245] In some embodiments, the targeting molecule comprises the peptide Ac EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9).

[00246] In some embodiments, the targeting molecule comprises the peptide Ac PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10).

[00247] In some embodiments, the targeting molecule comprises the peptide Ac SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11).

[00248] In some embodiments, the targeting molecule comprises the peptide Ac SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12).

[00249] In some embodiments, the targeting molecule comprises the peptide Ac SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13).

[00250] In some embodiments, the targeting molecule comprises the peptide Ac SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14).

[00251] In some embodiments, the targeting molecule comprises the peptide DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16).

[00252] In some embodiments, the targeting molecule comprises the peptide QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20).

[00253] In some embodiments, the targeting molecule comprises the peptide QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).

[00254] In some embodiments, the targeting molecule comprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22).

[00255] In some embodiments, the targeting molecule comprises the peptide EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23).

[00256] In some embodiments, the targeting molecule comprises the peptide PYYVVKKSS (HNP401-N-8; SEQ ID NO:24).

[00257] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25).

[00258] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26).

[00259] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27).

[00260] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEP (HNP401-C-8; SEQ ID NO:28).

[00261] In some embodiments, the targeting molecule comprises the peptide PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118).

[00262] In some embodiments, the targeting molecule comprises the peptide EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119).

[00263] In some embodiments, the targeting molecule comprises the peptide PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120).

[00264] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121).

[00265] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122).

[00266] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123).

[00267] In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124).

[00268] In some embodiments, the targeting molecule comprises the peptide 5FAM QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

[00269] In some embodiments, the targeting molecule comprises a peptide sequence sharing at least 80% homology with a peptide sequence disclosed herein. In some embodiments, the

targeting molecule comprises a peptide sequence sharing at least 85% homology with a peptide

sequence disclosed herein. In some embodiments, the targeting molecule comprises a peptide

sequence sharing at least 90% homology with a peptide sequence disclosed herein. In some

embodiments, the targeting molecule comprises a peptide sequence sharing at least 95% homology

with a peptide sequence disclosed herein. In some embodiments, the targeting molecule comprises

a peptide sequence sharing at least 99% homology with a peptide sequence disclosed herein.

[00270] In some embodiments, the targeting molecule comprises a peptide sequence having at least 75%, 80%, 85%, 90%, 95%, 97%, or 99% identity with a peptide sequence of

(HNP402 with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID

NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac

SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),

QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSSGG (HNP401-N-4; SEQ ID NO:22); EEPYYVVKKSS (HNP401-N

6; SEQ ID NO:23); PYYVVKKSS (HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ

ID NO:25), SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQ ID

SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),

ID NO:104).

[00271] In some embodiments, the targeting molecule comprises an aptamer.

[00272] The peptides and aptamers of the present invention are synthesized by any suitable method. For example, targeting peptides and aptamers of the present invention can be chemically

synthesized by solid phase peptide synthesis. In some embodiments, peptides of the present

invention are acetylated at the N-terminus ("Ac" or "acetyl"), amidated at the C-terminus ("CONH 2

" or "NH 2 "), or both. For example, the targeting peptide may comprise Ac-SGQVPWEEPYYVVKKSS (HNP

401; SEQ ID NO:43), Ac-WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:44), Ac-DLPDIIWDFNWETA

(HNP 403; SEQ ID NO:45), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac

WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC

(HNP403 with GGC linker; SEQ ID NO:6), Ac-DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:46), Ac

QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:47), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with

GGC linker; SEQ ID NO:7), Ac-QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:48),

Ac-PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:49), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC

linker; SEQ ID NO:8), Ac-EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:50), Ac-EEPYYVVKKSSGGC (HNP401

N-6 with GGC linker; SEQ ID NO:9), Ac- PYYVVKKSS (HNP401-N-8; SEQ ID NO:51), Ac-PYYVVKKSSGGC

(HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac-SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:52),

Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac- SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:53), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID

NO:12), Ac-SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:54), Ac-SGQVPWEEPYYGGC (HNP401-C-6 with

GGC linker; SEQ ID NO:13), Ac-SGQVPWEEP (HNP401-C-8; SEQ ID NO:55), Ac-SGQVPWEEPGGC

(HNP401-C-8 with GGC linker; SEQ ID NO:14), SGQVPWEEPYYVVKKSS-CONH 2 (HNP 401; SEQ ID

NO:56), WEYHYVDLNWTSQHPQ-CONH 2 (HNP 402; SEQ ID NO:57), DLPDIIWDFNWETA-CONH 2 (HNP

403; SEQ ID NO:58), SGQVPWEEPYYVVKKSSGGC-CONH 2 (HNP401 with GGC linker; SEQ ID NO:59),

WEYHYVDLNWTSQHPQGGC-CONH 2 (HNP402 with GGC linker; SEQ ID NO:60),

DLPDIIWDFNWETAGGC-CONH 2 (HNP403 with GGC linker; SEQ ID NO:61), DTHAHAKPRVPAFKSV

CONH 2 (HNP 404; SEQ ID NO:62), QVPWEEPYYVVKKSSGGC-CONH 2 (HNP401-N-2 with GGC linker; SEQ

ID NO:63), QVPWEEPYYVVKKSSGG-CONH 2 (HNP401-N-2 with GG linker; SEQ ID NO:64),

PWEEPYYVVKKSSGGC-CONH 2 (HNP401-N-4 with GGC linker; SEQ ID NO:65), EEPYYVVKKSSGGC

CONH 2 (HNP401-N-6 with GGC linker; SEQ ID NO:66), PYYVVKKSSGGC-CONH 2 (HNP401-N-8 with GGC

linker; SEQ ID NO:67), SGQVPWEEPYYVVKKGGC-CONH 2 (HNP401-C-2 with GGC linker; SEQ ID NO:68), SGQVPWEEPYYVVGGC-CONH 2 (HNP401-C-4 with GGC linker; SEQ ID NO:69), SGQVPWEEPYYGGC

CONH 2 (HNP401-C-6 with GGC linker; SEQ ID NO:70), SGQVPWEEPGGC-CONH 2 (HNP401-C-8 with

GGC linker; SEQ ID NO:71), QVPWEEPYYVVKKSS-CONH 2 (HNP401-N-2; SEQ ID NO:72),

PWEEPYYVVKKSS-CONH 2 (HNP401-N-4; SEQ ID NO:73), EEPYYVVKKSS-CONH 2 (HNP401-N-6; SEQ ID

NO:74), PYYVVKKSS-CONH 2 (HNP401-N-8; SEQ ID NO:75), SGQVPWEEPYYVVKK-CONH 2 (HNP401-C-2;

SEQ ID NO:76), SGQVPWEEPYYVV-CONH 2 (HNP401-C-4; SEQ ID NO:77), SGQVPWEEPYY-CONH 2

(HNP401-C-6; SEQ ID NO:78), and SGQVPWEEP-CONH 2 (HNP401-C-8; SEQ ID NO:79), Ac

SGQVPWEEPYYVVKKSS-CONH 2 (HNP401; SEQ ID NO:80), Ac-WEYHYVDLNWTSQHPQ-CONH 2

(HNP402; SEQ ID NO:81), Ac-DLPDIIWDFNWETA-CONH 2 (HNP403; SEQ ID NO:82), Ac

SGQVPWEEPYYVVKKSSGGC-CONH 2 (HNP401 with GGC linker; SEQ ID NO:83), Ac

WEYHYVDLNWTSQHPQGGC-CONH 2 (HNP402 with GGC linker; SEQ ID NO:84), Ac

DLPDIIWDFNWETAGGC-CONH 2 (HNP403 with GGC linker; SEQ ID NO:85), Ac-DTHAHAKPRVPAFKSV

CONH 2 (HNP 404; SEQ ID NO:86), Ac-QVPWEEPYYVVKKSSGGC-CONH 2 (HNP401-N-2 with GGC linker; SEQ ID NO:87), Ac-QVPWEEPYYVVKKSSGG-CONH 2 (HNP401-N-2 with GG linker; SEQ ID NO:88), Ac

PWEEPYYVVKKSSGGC-CONH 2 (HNP401-N-4 with GGC linker; SEQ ID NO:89), Ac-EEPYYVVKKSSGGC

CONH 2 (HNP401-N-6 with GGC linker; SEQ ID NO:90), Ac-PYYVVKKSSGGC-CONH 2 (HNP401-N-8 with

GGC linker; SEQ ID NO:91), Ac-SGQVPWEEPYYVVKKGGC-CONH 2 (HNP401-C-2 with GGC linker; SEQ ID

NO:92), Ac-SGQVPWEEPYYVVGGC-CONH 2 (HNP401-C-4 with GGC linker; SEQ ID NO:93), Ac

SGQVPWEEPYYGGC-CONH2 (HNP401-C-6 with GGC linker; SEQ ID NO:94), Ac-SGQVPWEEPGGC

CONH 2 (HNP401-C-8 with GGC linker; SEQ ID NO:95), Ac-QVPWEEPYYVVKKSS-CONH 2 (HNP401-N-2;

SEQ ID NO:96), Ac-PWEEPYYVVKKSS-CONH 2 (HNP401-N-4; SEQ ID NO:97), Ac-EEPYYVVKKSS-CONH 2

(HNP401-N-6; SEQ ID NO:98), Ac-PYYVVKKSS-CONH 2 (HNP401-N-8; SEQ ID NO:99), Ac SGQVPWEEPYYVVKK-CONH 2 (HNP401-C-2; SEQ ID NO:100), Ac-SGQVPWEEPYYVV-CONH 2 (HNP401-C

4; SEQ ID NO:101), Ac-SGQVPWEEPYY-CONH 2 (HNP401-C-6; SEQ ID NO:102), or Ac-SGQVPWEEP

CONH 2 (HNP401-C-8; SEQ ID NO:103). Techniques for solid phase synthesis are described, for

example, by Barany and Merrifteld (1963) Solid-Phase Peptide Synthesis; pp. 3-284 in The Peptides:

Analysis, Synthesis, Biology. Vol. 2: Special Methods in Peptide Synthesis, Part A.; Merrifield et al.

(1963) J. Am. Chem. Soc, 85: 2149-2156, and Stewart et al. (1984) Solid Phase Peptide Synthesis, 2nd

ed. Pierce Chem. Co., Rockford, 111.

V. Cargo

[00273] In some embodiments, the human neuron or nerve targeting molecule further comprises a cargo. In some embodiments, a targeting peptide comprises an amino acid sequence of

NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac

Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker;

SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),

SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),

ID NO:104).

[00274] In some embodiments, the human neuron or nerve targeting molecule further comprises a cargo. In some embodiments, the human neuron or nerve targeting molecule comprises

a peptide sequence selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID

DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac

DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20),

SEQ ID NO:22), EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS (HNP401-N-8; SEQ ID NO:24),

SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26),

SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28),

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122), SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), and 5FAM

QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104). In some embodiments

the targeting molecule comprises a peptide selected from the group consisting of

and DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3). In some embodiments, the targeting molecule

comprises a peptide selected from the group consisting of SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID

NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac

QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), SGQVPWEEPYYVVKK (HNP401

C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG

(HNP401-N-2 with GG linker; SEQ ID NO:21), and 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2

with GG linker; SEQ ID NO:104). In some embodiments, the targeting molecule comprises the

peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In some embodiments, the targeting molecule comprises the peptide WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In some

embodiments, the targeting molecule comprises the peptide DLPDIIWDFNWETA (HNP 403; SEQ ID

NO:3). In some embodiments, the targeting molecule comprises the peptide Ac

SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4). In some embodiments, the

targeting molecule comprises the peptide Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker;

SEQ ID NO:5). In some embodiments, the targeting molecule comprises the peptide Ac

DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6). In some embodiments, the

targeting molecule comprises the peptide QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some

embodiments, the targeting molecule comprises the peptide QVPWEEPYYVVKKSSGG (HNP401-N-2

with GG linker; SEQ ID NO:21). In some embodiments, the targeting molecule comprises the peptide

Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7). In some embodiments, the

targeting molecule comprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22). In some

embodiments, the targeting molecule comprises the peptide Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8). In some embodiments, the targeting molecule comprises the peptide

EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23). In some embodiments, the targeting molecule

comprises the peptide Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9). In some

embodiments, the targeting molecule comprises the peptide PYYVVKKSS (HNP401-N-8; SEQ ID

NO:24). In some embodiments, the targeting molecule comprises the peptide Ac-PYYVVKKSSGGC

(HNP401-N-8 with GGC linker; SEQ ID NO:10). In some embodiments, the targeting molecule

comprises the peptide SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25). In some embodiments, the

targeting molecule comprises the peptide Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11). In some embodiments, the targeting molecule comprises the peptide

SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26). In some embodiments, the targeting molecule

comprises the peptide Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12). In

some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYY (HNP401-C-6;

SEQ ID NO:27). In some embodiments, the targeting molecule comprises the peptide Ac

SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13). In some embodiments, the

targeting molecule comprises the peptide SGQVPWEEP (HNP401-C-8; SEQ ID NO:28). In some

embodiments, the targeting molecule comprises the peptide 5FAM-QVPWEEPYYVVKKSSGG-NH2

(HNP401-N-2 with GG linker; SEQ ID NO:104). In some embodiments, the targeting molecule

comprises the peptide Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14). In some

embodiments, the targeting molecule comprises the peptide DTHAHAKPRVPAFKSV (HNP 404; SEQ ID

NO:16). In some embodiments, the targeting molecule comprises a peptide that is not Ac

SHSNTQTLAKAPEHTGC (Ac-NP41 with GC linker; SEQ ID NO:17). In some embodiments, the targeting molecule comprises a peptide that is not SHSNTQTLAKAPEHTGC (NP41 with GC linker; SEQ ID

NO:18). In some embodiments, the peptide is not NTQTLAKAPEHT (NP41; SEQ ID NO:19).

[00275] In some embodiments, the peptide or aptamer is directly bound to a cargo. In some embodiments, the peptide or aptamer is indirectly (e.g., via a linker) bound to a cargo. In some

embodiments, the peptide or aptamer is bound to a cargo at its N-terminus, at its C-terminus, or at an internal position (e.g., to an internal amino acid) of the peptide or aptamer. In some

embodiments, two, three, four or more peptides or aptamers are directly or indirectly bound to a

cargo. In certain embodiments, a cargo is a drug, fluorescent moeity, photosensitizing agent, or a

combination thereof. In some embodiments, the cargo is a drug. In some embodiments, the cargo is

a fluorescent moiety or a fluorescent dye. In some embodiments, the cargo comprises a fluorescent

moiety or a fluorescent dye. In some embodiments, the cargo is a photosensitizing agent. In some

embodiments, the peptide or aptamer is bound to two or more cargo moieties. The two or more

cargo moieties may be the same moiety or different moieties, or be from the same class of cargo

moieties (e.g., two drugs) or from different classes of cargo moieties (e.g., one drug and one

fluorescent moiety).

[00276] Common classes of fluorescent dyes include, but are not limited to, xanthenes such as rhodamines, rhodols and fluoresceins, and their derivatives; bimanes; coumarins and their derivatives such as umbelliferone and aminomethyl coumarins; aromatic amines such asdansyl; squarate dyes; benzofurans; fluorescent cyanines; carbazoles; dicyanomethylene pyranes, polymethine, oxabenzanthrane, xanthene, pyrylium, carbostyl, perylene, acridone, quinacridone, rubrene, anthracene, coronene, phenanthrecene, pyrene, butadiene, stilbene, lanthanide metal chelate complexes, rare-earth metal chelate complexes, and derivatives of such dyes. Fluorescent dyes are discussed, for example, in U.S. Pat. No. 4,452,720; U.S. Pat. No. 5,227,487; and U.S. Pat. No. 5,543,295.

[00277] In some embodiments, the fluorescent moiety or dye selected from the group consisting of a xanthene; a bimane; a coumarin; an aromatic amines; a benzofuran; a fluorescent cyanine; a carbazole; a dicyanomethylene pyrane; polymethine; oxabenzanthrane; pyrylium; carbostyl; perylene; acridone; quinacridone; rubrene; anthracene; coronene; phenanthrecene; pyrene; butadiene; stilbene; porphyrin; pthalocyanine; lanthanide metal chelate complexes; rare earth metal chelate complexes; FITC; Cy3; EGFP; cyan fluorescent protein (CFP); EGFP; 5-FAM; 6 FAM; FAM; fluorescein, IAEDANS, EDANS and BODIPY FL; TRITC; Cy5; Cy3; YFP; 6-FAM; LC Red 640; Alexa Fluor 546; fluorescein; tetramethylrhodamine; Dabcyl; BODIPY FL; QSY 7, QSY 9, QSY 21 and BBQ-650 dyes.

[00278] In some embodiments, the cargo comprises fluorescein dyes. Typical fluorescein dyes include, but are not limited to, 5-carboxyfluorescein, fluorescein-5-isothiocyanate, 5(6) carboxyfluorescein, 5,6-dicarboxyfluorescein, 5-(and 6)-sulfofluorescein, sulfonefluorescein, succinyl fluorescein, 5-(and 6)-carboxy SNARF-1, carboxyfluorescein sulfonate, carboxyfluorescein zwitterion, carboxyfluorescein quaternary ammonium, carboxyfluorescein phosphonate, carboxyfluorescein GABA, carboxyfluorescein-cys-Cy5, 5'(6')-carboxyfluorescein, fluorescein glutathione, and 6 carboxyfluorescein; examples of other fluorescein dyes can be found, for example, in U.S. Pat. No. 6,008,379, U.S. Pat. No. 5,750,409, U.S. Pat. No. 5,066,580, and U.S. Pat. No. 4,439,356. A cargo may include a rhodamine dye, such as, for example, 5-(and 6)-carboxy rhodamine 110, tetramethylrhodamine-6-isothiocyanate, 5-carboxytetramethylrhodamine, 5-carboxy rhodol derivatives, tetramethyl and tetraethyl rhodamine, diphenyldimethyl and diphenyldiethyl rhodamine, dinaphthyl rhodamine, rhodamine 101 sulfonyl chloride (sold under the tradename of TEXAS RED®), and other rhodamine dyes. Other rhodamine dyes can be found, for example, in U.S. Pat. No. 6,080,852; U.S. Pat. No. 6,025,505; U.S. Pat. No. 5,936,087; U.S. Pat. No. 5,750,409. In some embodiments, a cargo moiety includes a cyanine dye, such as, for example, Cy3, Cy3B, Cy3.5,

Cy5, Cy5.5, Cy 7.

[00279] In some embodiments, cargo moiety comprises fluorophores. Fluorophores are commercially available and any known and/or commercially available fluorophore can be employed

as the cargo. In some embodiments, the fluorophore exhibits green fluorescence (such as for

example 494 nm/519 nm), orange fluorescence (such as for example 554 nm/570 nm), red

fluorescence (such as for example 590 nm/617 nm), or far red fluorescence (such as for example 651

nm/672 nm) excitation/emission spectra. In some embodiments, the fluorophore is a fluorophore with excitation and emission spectra in the range of about 350 nm to about 775 nm. In some

embodiments the excitation and emission spectra are about 346 nm/446 nm, about 494 nm/519

nm, about 554 nm/570 nm, about 555 nm/572 nm, about 590 nm/617 nm, about 651 nm/672 nm,

about 679 nm/702 nm or about 749 nm/775 nm. In some embodiments, the fluorophore can

include but is not limited to AlexaFluor 3, AlexaFluor 5, AlexaFluor 350, AlexaFluor 405, AlexaFluor

430, AlexaFluor 488, AlexaFluor 500, AlexaFluor 514, AlexaFluor 532, AlexaFluor 546, AlexaFluor 555,

AlexaFluor 568, AlexaFluor 594, AlexaFluor 610, AlexaFluor 633, AlexaFluor 647, AlexaFluor 660,

AlexaFluor 680, AlexaFluor 700, and AlexaFluor 750 (Molecular Probes AlexaFluor dyes, available

from Life Technologies, Inc. (USA)). In some embodiments, the fluorophore can include but is not

limited to Cy dyes, including Cy2, Cy3, Cy3B, Cy3.5, Cy5, Cy5.5 and Cy7 (available from GE Life

Sciences or Lumiprobes). In some embodiments the fluorophore can include but is not limited to

DyLight 350, DyLight 405, DyLight 488, DyLight 550, DyLight 594, DyLight 633, DyLight 650, DyLight

680, DyLight 750 and DyLight 800 (available from Thermo Scientific (USA)). In some embodiments, the fluorophore can include but is not limited to a FluoProbes 390, FluoProbes 488, FluoProbes 532,

FluoProbes 547H, FluoProbes 594, FluoProbes 647H, FluoProbes 682, FluoProbes 752 and

FluoProbes 782, AMCA, DEAC (7-Diethylaminocoumarin-3-carboxylic acid); 7-Hydroxy-4

methylcoumarin-3; 7-Hydroxycoumarin-3; MCA (7-Methoxycoumarin-4-acetic acid); 7

Methoxycoumarin-3; AMF (4'-(Aminomethyl)fluorescein); 5-DTAF (5-(4,6

Dichlorotriazinyl)aminofluorescein); 6-DTAF (6-(4,6-Dichlorotriazinyl)aminofluorescein); FAM; 6-FAM

(6-Carboxyfluorescein), 5(6)-FAM cadaverine; 5-FAM cadaverine; 5(6)-FAM ethylenediamme; 5-FAM

ethylenediamme; 5-FITC (FITC Isomer I; fluorescein-5-isothiocyanate); 5-FITC cadaverin; Fluorescein

5-maleimide; 5-IAF (5-lodoacetamidofluorescein); 6-JOE (6-Carboxy-4',5'-dichloro-2',7'

dimethoxyfluorescein); 5-CRI 10 (5-Carboxyrhodamine 110); 6-CRI 10 (6-Carboxyrhodamine 110); 5

CR6G (5-Carboxyrhodamine 6G); 6-CR6G (6-Carboxyrhodamine 6G); 5(6)-Carboxyrhodamine 6G

cadaverine; 5(6)-Caroxyrhodamine 6G ethylenediamme; 5-ROX (5-Carboxy-X-rhodamine); 6-ROX (6

Carboxy-X-rhodamine); 5-TAMRA (5-Carboxytetramethylrhodamine); 6-TAMRA (6

Carboxytetramethylrhodamine); 5-TAMRA cadaverine; 6-TAMRA cadaverine; 5-TAMRA

ethylenediamme; 6-TAMRA ethylenediamme; 5-TMR C6 maleimide; 6-TMR C6 maleimide; TR C2 maleimide; TR cadaverine; 5-TRITC; G isomer (Tetramethylrhodamine-5-isothiocyanate); 6-TRITC; R

isomer (Tetramethylrhodamine-6-isothiocyanate); Dansyl cadaverine (5

Dimethylaminonaphthalene-l-(N-(5-aminopentyl))sulfonamide); EDANS C2 maleimide;

fluorescamine; NBD; and pyrromethene and derivatives thereof.

[00280] In some embodiments, a cargo comprises an environmentally sensitive fluorescent dye or fluorophore. Examples of environmentally sensitive fluorescent dyes or fluorophores include

5,6-carboxy-diethyl rhodol (pH sensitive), merocyanine (membrane potential sensitive), and Nile red

carboxylic acid (lipid sensitive).

[00281] In some embodiments, a cargo comprises a photosensitizing agent. A photosensitizing agent is any agent or compound useful in light induced ablation therapy. Such

agents, when exposed to a specific wavelength of light, react with molecular oxygen to produce

singlet oxygen, which is highly cytotoxic. Thus, targeting molecules of the present invention

comprising a photosensitizing agent may be used to focally injure nerves. In certain embodiments, a

photosensitizing agent is a porphyrin, chlorin, or dye. Examples of photosensitizing agents include

porphyrin, protoporfin IX, purlytin, verteporfin, HPPH, temoporfin, methylene blue, photofrin,

protofrin, hematoporphyrin, Talaporfin, benzopophyrin derivative monoacid, 5-aminileuvolinic acid,

Lutetium texaphyrin, metallophthalocyanine, metallo-naphthocyaninesulfobenzo-porphyrazines,

metallo-naphthalocyanines, zinc tetrasulfophthalocyanine, bacteriochlorins, metallochlorins,

chlorine derivative, Tetra(m-hydroxyphenyl)chlorin (mTHPC), pheophorbide, dibromofluorescein

(DBF), IR700DX, naphthalocyanine, and porphyrin derivatives. In some embodiments, the

photosensitizing agent is conjugated to a C-terminal cysteine residue of the human neuron or nerve

targeting molecule via maleimide mediated conjugation. Preferably, the photosensitizing agent of

the present invention is activated by light having a wavelength of between 400 nm to 700 nm. Still

more preferably, the photosensitizing agent in the present invention is activated at 627 nm and 660

nm. An optimal light dose can be identified to generate maximal nerve killing with minimal injury to

adjacent tissue.

VI. Drugs

[00282] In some embodiments, the human neuron or nerve targeting molecule further comprises a drug. All drugs that act on a neuron or nerve (or a component thereof) are

encompassed within the term "drug." Specific examples of drug given herein, are illustrative and are

not meant to limit the drugs for use with the targeting molecules disclosed herein. In some

embodiments, the peptide or aptamer is directly bound to a drug. In some embodiments, the

peptide or aptamer is indirectly (e.g., via a linker) bound to a drug. In some embodiments, two or

more peptides or aptamers are directly or indirectly bound to a drug. In some embodiments, the human neuron or nerve targeting molecule further comprises a cargo. In some embodiments, the

human neuron or nerve targeting molecule comprises a peptide sequence selected from the group

NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac

WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC

SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac

NO:11), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4;

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), or 5FAM

the targeting molecule comprises a peptide selected from the group consisting of:

SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3). In some embodiments, the targeting molecule comprises a peptide selected from the group consisting of SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID

NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), SGQVPWEEPYYVVKK (HNP401

peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In some embodiments, the targeting

molecule comprises the peptide WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In some

NO:3). In some embodiments, the targeting molecule comprises the peptide Ac

targeting molecule comprises the peptide QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some embodiments, the targeting molecule comprises the peptide QVPWEEPYYVVKKSSGG (HNP401-N-2

embodiments, the targeting molecule comprises the peptide Ac-PWEEPYYVVKKSSGGC (HNP401-N-4

with GGC linker; SEQ ID NO:8). In some embodiments, the targeting molecule comprises the peptide

embodiments, the targeting molecule comprises the peptide Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14). In some embodiments, the targeting molecule comprises the peptide

DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16). In some embodiments, the targeting molecule

comprises the peptide 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID

NO:104).

[00283] In some embodiments, the drug is selected from a drug that: induces cell death (apoptotic or necrotic), inhibits cell death (apoptotic or necrotic), inhibits the transmission of a

neuron or nerve signal (i.e., an electrochemical impulse), inhibits the release of a neurotransmitter,

agonizes the activity of a GABA receptor, partially or fully inhibits the repolarization of a neuron,

disrupts the conduction of an ion channel, or a combination thereof.

[00284] In some embodiments, the drug is an antihistamine, a GABA receptor modulator, a neurotransmitter reuptake inhibitor, a local anesthetic, an anticholinergic, a sodium channel blocker,

a calcium channel blocker, a thyrotropin-releasing hormone, ay-secretase inhibitor, an AMPA

receptor agonist or antagonist, an NMDA receptor agonist or antagonist, an mGlu receptor agonist

or antagonist, a growth factor, an antiemetic agent, a corticosteroid; a cytotoxic agent; an

antioxidant, an iron chelator, a mitochondrial modulator, a sirtuin modulator, a nitric oxide (NO)

and/or nitric oxide synthase (NOS) modulator, a potassium channel agonist or antagonist, a

purigenic receptor agonist or antagonist, or a combination thereof.

[00285] In some embodiments, the drug is meclizine, diphenhydramine, dimenhydrinate, loratadine, quetiapine, mepyramine, piperoxan, antazoline, carbinoxamine, doxylamine, clemastine,

pheniramine, chlorphenamine, chlorpheniramine, dexchlorpheniramine, brompheniramine,

triprolidine, cyclizine, chlorcyclizine, hydroxyzine, promethazine, alimemazine, trimeprazine,

cyproheptadine, azatadine, ketotifen, oxatomide, meclizine hydrochloride, promethazine

hydrochloride, cinnarizine, hydroxyzine pamoate, betahistine dihydrochloride, alprazolam,

bromazepam, brotizolam, chlordiazepoxide, clonazepam, clorazepate, diazepam, estazolam,

flunitrazepam, flurazepam, loprazolam, lorazepam, lormetazepam, idazolam, nimetazepam,

nitrazepam, oxazepam, prazepam, temazepam, triazolam, clonazepam, diazepam, lorazepam,

furosemide, bumetanide, ethacrynic acid, gabapentin, pregabalin, muscimol, baclofen, amitriptyline, nortriptyline, trimipramine, fluoxetine, paroxetine, sertraline, glycopyrrolate, homatropine, scopolamine, atropine, benzocaine, carticaine, cinchocaine, cyclomethycaine, lidocaine, prilocaine, propxycaine, proparacaine, tetracaine, tocainide, trimecaine, carbamazepine, oxcarbazepine, phenytein, valproic acid, sodium valproate, cinnarizine, flunarizine, nimodipine, thyrotropin releasing hormone, amifostine (also known as WR-2721, or ETHYOL*); a carbamate compound (e.g.,

2-phenyl-1,2-ethanediol monocarbomates and dicarbamates); LY450139 (hydroxylvaleryl

monobenzocaprolactam); L685458 (IS-benzyl-4R[1-[1-S-carbamoyl-2-phenethylcarbamoyl)- IS-3

methylbutylcarbamoyl]-2R-hydroxy-5-phenylpentyl}carbamic acid tert-butyl ester); LY411575 (N2

[(2S)-2-(3,5-difluorophenyl)-2-hydroxyethanoyl]-N1[(7S)-5-methyl-6-oxo-6,7-dihydro-5H

dibenzo[bid]azepin-7yl]-L-alaninamide); MK-0752; tarenflurbil; BMS-299897 (2-[(IR)--[[(4

chlorophenyl) sulfony](2,5-difluorophenyl)amino]ethyl]-5-fluorobenzenepropanoic acid; CNQX (6

cyano-7-nitroquinoxaline-2,3-dione); NBQX (2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo[f]quinoxaline

2,3-dione); DNQX (6,7-dinitroquinoxaline-2,3-dione); kynurenic acid; 2,3-dihydroxy-6-nitro-7

sulfamoylbenzo-[fjquinoxaline; 1-aminoadamantane; dextromethorphan; dextrorphan; ibogaine;

ketamine; nitrous oxide; phencyclidine; riluzole; tiletamine; memantine;dizocilpine; aptiganel;

remacimide; 7-chlorokynurenate; DCKA (5,7-dichlorokynurenic acid); kynurenic acid; 1

aminocyclopropanecarboxylic acid (ACPC); AP7 (2-amino-7-phosphonoheptanoic acid); APV (R-2 amino-5-phosphonopentanoate); CPPene (3-[(R)-2-carboxypiperazin-4-yl]-prop-2-enyl--phosphonic

acid); (+)-(IS, 2S)--(4-hydroxy-phenyl)-2-(4-hydroxy-4-phenylpiperidino)- 1-pro-panol; ( 1 S, 2S)- 1

(4-hydroxy-3-methoxyphenyl)-2-(4-hydroxy-4-phenylpiperi-dino)- 1 -propanol; (3R, 4S)-3-(4-(4

fluorophenyl)-4-hydroxypiperidin-1-yl-)-chroman-4;7-diol;(IR*, 2R*)--(4-hydroxy-3-memylphenyl)

2-(4-(4-fluoro-phenyl)-4-hydroxypiperidin- 1 -yl)-propan- 1 -ol-mesylate); LY389795 ((-)-2-thia-4

aminobicyclo-hexane-4,6-dicarboxylate); LY379268 ((-)-2-oxa-4-aminobicyclo-hexane-4,6

dicarboxylate); LY354740 ((+)-2-aminobicyclo-hexane-2,6dicarboxylate); DCG-IV ((2S,2'R,3'R)-2

(2',31-dicarboxycyclopropyl)glycine); 2R,4R-APDC (2R,4R-4-aminopyrrolidine-2,4-dicarboxylate); (S)

3C4HPG ((S)-3-carboxy-4-hydroxyphenylglycine); (S)-4C3HPG ((S)-4-carboxy-3

hydroxyphenylglycine); L-CCG-1 ((2S, 1'S,2'S)-2-(carboxycyclopropyl)glycine); ACPT-1 ((IS,3R,4S)-

aminocyclopentane-1,3,4-tricarboxylic acid); L-AP4 (L-(+)-2-Amino-4-phosphonobutyric acid); (S)-3,4

DCPG ((S)-3,4-dicarboxyphenylglycine); (RS)-3,4-DCPG ((RS)-3,4-dicarboxyphenylglycine); (RS)-4

phosphonophenylglycine ((RS)PPG); AMN082 (,N'-bis(diphenylmethyl)-1,2-ethanediamine dihydrochloride); DCG-IV ((2S,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl)glycine); AMN082; brain-derived

neurotrophic factor (BDNF); ciliary neurotrophic factor (CNTF); glial cell-line derived neurotrophic

factor (GDNF); neurotrophin-3; neurotrophin-4; fibroblast growth factor (FGF) receptor; insulin-like

growth factor (IGF); an aminoglycoside antibiotic (e.g., gentamicin and amikacin); a macrolide

antibiotic (e.g, erythromycin); a glycopeptide antibiotic (e.g. vancomycin); salicylic acid; nicotine;

Eburnamenine-14-carboxylic acid ethyl ester; sipatrigine (2-(4-Methylpiperazin-1-yl)-5-(2,3,5

trichlorophenyl)-pyrimidin-4-amine); amiloride (3,5-diamino-N- (aminoiminomethyl)-6

chloropyrazinecarbox amide hydrochloride); carbamazepine (5H-dibenzo[b,f]azepine-5 carboxamide); TTX (octahydro-12-(hydroxymethyl)-2-imino- 5,9:7,10a-dimethan o-OaH

[1,3]dioxocino[6,5-d]pyrimidine-4,7,10,II,12-pen tol); RS100642 (I-(2,6-dimethyl-phenoxy)-2

ethylaminopropane hydrochloride); mexiletine ((1- (2,6-dimethylphenoxy)-2-aminopropane

hydrochloride)); QX-314 (N-(2,6- Dimethylphenylcarbamoylmethyl)triethylammonium bromide);

phenytoin (5,5-diphenylimidazolidine-2,4-dione); lamotrigine (6-(2,3-dichlorophenyl)-1,2,4-triazine

3,5-diamine); 4030W92 (2,4-diamino-5-(2,3-dichlorophenyl)-6-fluoromethylpyrimidine); BW1003C87

(5-(2,3,5-trichlorophenyl) pyrimidine-2,4- 1.1 ethanesulphonate); QX-222 (2- [(2,6

dimethylphenyl)amino]-N,N,N-trimethyl-2-oxoetha niminium chloride); ambroxol (trans-4- [ [(2

Amino-3, 5 -dibromophenyl)methyl] amino] cyclo hexanol hydrochloride) ; R56865 (N-[ 1 -(4-(4

fluo rophenoxy) butyl]-4-piperidinyl-N-methyl-2-benzo-thiazolamine); lubeluzole; ajmaline

((17R,21alpha)-ajmalan-17,21-diol); procainamide (4-amno-N-(2-diethylaminoethyljbenzamide

hydrochloride); flecainide; riluzoleor; triamicinolone actenoide; Dexamethasone; promethazine;

prochlorperazine; trimethobenzamide; triethylperazine; dolasetron; granisetron; ondansetron; tropisetron; and palonosetron; droperidol; meclizine; perphenazine; thiethyl perazine;

domperidone; properidol; haloperidol; chlorpromazine; promethazine; prochlorperazine;

metoclopramide; dronabinol; nabilone; sativex; scopolamine; dexamethasone; trimethobenzamine;

emetrol; propofol; muscimol; acridine carboxamide; actinomycin; 17-N-allylamino-17

demethoxygeldanamycin; amsacrine; aminopterin; anthracycline; antineoplastic; antineoplaston; 5

azacytidine; azathioprine; BL22; bendamustine; biricodar; bleomycin; bortezomib; bryostatin;

busulfan; calyculin; camptothecin; capecitabine; carboplatin; chlorambucil; cisplatin; cladribine;

clofarabine; cytarabine; dacarbazine; dasatinib; daunorubicin; decitabine; dichloroacetic acid;

discodermolide; docetaxel; doxorubicin; epirubicin; epothilone; eribulin; estramustine; etoposide;

exatecan; exisulind; ferruginol; floxuridine; fludarabine; fluorouracil; fosfestrol; fotemustine;

gemcitabine; hydroxyurea; IT-101; idarubicin; ifosfamide; imiquimod; irinotecan; irofulven;

ixabepilone; laniquidar; lapatinib; lenalidomide; lomustine; lurtotecan; mafosfamide; masoprocol;

mechlorethamine; melphalan; mercaptopurine; mitomycin; mitotane; mitoxantrone; nelarabine; nilotinib; oblimersen; oxaliplatin; PAC-1; methotrexate (RHEUMATREX©, Amethopterin);

cyclophosphamide (CYTOXAN©) thalidomide (THALID OMID*); paclitaxel; pemetrexed; pentostatin;

pipobroman; pixantrone; plicamycin; procarbazine; proteasome inhibitors (e.g.; bortezomib);

raltitrexed; rebeccamycin; rubitecan; SN-38; salinosporamide A; satraplatin; streptozotocin;

swainsonine; tariquidar; taxane; tegafur-uracil; temozolomide; testolactone; thioTEPA; tioguanine; topotecan; trabectedin; tretinoin; triplatin tetranitrate; tris(2-chloroethyl)amme; troxacitabine; uracil mustard; valrubicin; vinblastine; vincristine; vinorelbine; vorinostat; zosuquidar; N acetylcysteine; vitamin E; vitamin C; vitamin A; lutein; selenium glutathione; melatonin; a polyphenol; a carotenoid; coenzyme Q-0; Ebselen (2-phenyl-1, 2-benzisoselenazol-3(2H)-one (also called PZ 51 or DR3305); L-methionine; azulenyl nitrones; L-(+)-Ergothioneine; CAPE (caffeic acid phenethyl ester); dimethylthiourea; dimethylsulfoxide; disufenton sodium; pentoxifylline; MCI-186;

Ambroxol; U-83836E; MitoQ (mitoquinone mesylate); Idebenone (2-(10-hydroxydecyl)-5,6

dimethoxy-3-methyl-cyclohexa-2,5-diene-1,4-dione); desferrioxamine; hydroxybenzyl ethylene

diamine; fullerenol-1, pyrrolidine dithiocarbamate; acetylcarnitine; lipoic acid; a stilbene; a chalcone;

a flavone; an isoflavone; a flavanones; an anthocyanidin; a catechin; isonicotinamide;dipyridamole;

ZM 336372; camptothecin; coumestrol; nordihydroguaiaretic acid; esculetin; SRT-1720; SRT-1460;

SRT-2183; aminoguanidine; I-Amino-2-hydroxyguanidine p-toluensulfate; GED; bromocriptine

mesylate; dexamethasone; SDMA; ADMA; L-NMMA; L- NMEA; D-MMA; L-NIL; L-NNA; L-NPA; L

NAME; L-VNIO; diphenyleneiodonium chloride; 2-ethyl-2-thiopseudourea; haloperidol; L-NIO; MEG;

SMT; SMTC; 7-Ni; nNOS inhibitor; 1,3-PBITU; L-thiocitrulline; TRIM; MTR-105; BBS-1; BBS-2; ONO

1714; GW273629; GW 274150; PPA250; AR-R17477; AR-R18512; spiroquinazolone; 1400W; S- NC; NTG; SNP; thapsigargin; VEGF; bradykinin; ATP; sphingosine-1 -phosphate; estrogen; angiopoietin;

acetylcholine; SIN-1; GEA 3162; GEA; GEA 5024; GEA 5538; SNAP; molsidomine; CNO-4; CNO-5;

DEA/NO; IPA/NO; SPER/NO; SULFI/NO; OXI/NO; DETA/NO; nicorandil; minoxidil, levcromakalim;

lemakalim; cromakalim; L-735,334; retigabine; flupirtine; BMS-204352; DMP-543; linopirdine; XE991;

4- AP; 3,4-DAP; E-4031; DIDS; Way 123,398; CGS- 12066 A; dofetilide; sotalol; apamin; amiodarone;

azimilide; bretylium; clofilium; tedisamil; ibutilide; sematilide; nifekalant; tamulustoxin; ATP; ADP;

UTP; UDP; UDP-glucose; adenosine; 2-MESATP; 2-MESADP; ABMEATP; DATPAS; ATPrS; BZ-ATP;

MRS2703; DENUFOSOL TETRASODIUM; MRS2365; MRS 2690; PSB 0474; A-317491; RO-3 (Roche);

SURAMIN; PPADS; PPNDS; DIDS; pyridoxal-5-phosphate; 5-(3-bromophenyl)-1,3-dihydro-2H

benzofuro- [3,2-e]-1,4-diazepin-2-one; cibacron blue; basilen blue; ivermectin; A-438079; A-740003;

NF023; NF449; NFI 10; NF157; MRS 2179; NF279; MRS 2211; MRS 2279; MRS 2500 tetrasodium salt;

TNP-ATP; tetramethylpyrazine; Ip5; jQy-carboxymethylene ATP; 3y-chlorophosphomethylene ATP;

KN-62; spinorphin; minocycline; SB-203580 (4-(4-Fluorophenyl)-2-(4-methylsulfmyl phenyl)-5-(4 pyridyl) IH-imidazole); PD 169316 (4-(4-Fluorophenyl)-2-(4-nitrophenyl)-5-(4-pyridyl)-H-imidazole);

SB 202190 (4-(4-Fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)lH-imidazole); RWJ 67657 (4-[4-(4

fluorophenyl)-l-(3-phenylpropyl)-5-(4-pyridinyl)-IH-imidazol -2-yl]-3-butyn-1-ol); SB 220025 (5-(2

Amino-4-pyrimidinyl)-4-(4-fluorophenyl)--(4-piperidinlyl)imidazole); D-JNKI-I ((D)-hJIPi75-i57-DPro

DPro-(D)-HIV-T AT57-48); AM-111 (Auris); SP600125 (anthra[1,9-cd]pyrazol-6(2H)-one); JNK Inhibitor

I ((L)-HIV-TAT48-57-PP-JBD20); JNK Inhibitor Ill ((L)-HIV-TAT47-57-gaba-c-Jun533-57); AS601245 (1,3

benzothiazol-2-yl (2-[[2-(3-pyridinyl) ethyl] amino]-4 pyrimidinyl) acetonitrile); JNK Inhibitor VI (H2N

RPKRPTTLNLF-NH2); JNK Inhibitor Vill (N-(4-Amino-5-cyano-6-ethoxypyridin-2-yl)-2-(2,5 dimethoxyphenyl)acetamide); JNK Inhibitor IX (N-(3-Cyano-4,5,6,7-tetrahydro--benzothien-2-yl)-l

naphthamide); dicumarol (3,3 '-Methylenebis(4-hydroxycoumarin)); SC-236 (4-[5-(4-chlorophenyl)-3

(trifluoromethyl)-IH-pyrazol-1-yljbenzene-sulfonamide); CEP-1347 (Cephalon); CEP-11004 (Cephalon);

an artificial protein comprising at least a portion of a Bc-2 polypeptide; a recombinant FNK; V5 (also

known as Bax inhibitor peptide V5); Bax channel blocker (()--(3,6-Dibromocarbazol-9-yl)-3

piperazin-1-yl-propan-2-ol); Bax inhibiting peptide P5 (also known as Bax inhibitor peptideP5); Kp7-6;

FAIM(S) (Fas apoptosis inhibitory molecule-short); FAIM(L) (Fas apoptosis inhibitory molecule-long);

Fas:Fc; FAP-1; NOK2; F2051; F1926; F2928; ZB4; Fas M3 mAb; EGF; 740 Y-P; SC 3036

(KKHTDDGYMPMSPGVA); PI 3-kinase Activator (Santa Cruz Biotechnology, Inc.); Pam3Cys ((S)-(2,3

bis(palmitoyloxy)-(2RS)-propyl)-N-palmitoyl-(R)-Cys-(S)-Ser(S)-Lys4-OH, trihydrochloride); Actl (NF-kB

activator 1); an anti-DcB antibody; Acetyl- 11-keto-b-Boswellic Acid; Andrographolide; Caffeic Acid

Phenethyl Ester (CAPE); Gliotoxin; Isohelenin; NEMO-Binding Domain Binding Peptide

(DRQIKIWFQNRRMKWKKTALD WSWLQTE); NF-kB Activation Inhibitor (6-Amino-4-(4 phenoxyphenylethylamino)quinazoline); NF-kB Activation Inhibitor II (4-Methyl-N-(3

phenylpropyl)benzene-1,2-diamine); NF-kB Activation Inhibitor Ill (3-Chloro-4-nitro-N-(5-nitro-2

thiazolyl)-benzamide); NF-kB Activation Inhibitor IV ((E)-2-Fluoro-4'-methoxystilbene); NF-kB

Activation Inhibitor V (5-Hydroxy-(2,6-diisopropylphenyl)- IH-isoindole-1,3-dione); NF-kB SN50 ( AAV

ALLP A VLLALL AP VQRKRQKLMP); Oridonin; Parthenolide; PPM-18 (2-Benzoylamino-1,4

naphthoquinone); Ro06-9920; Sulfasalazine; TIRAP Inhibitor Peptide (RQIKiWFNRRMKWKKLQLRD

AAPGGAIVS); Withaferin A; Wogonin; BAY 11-7082 ((E)3-[(4-Methylphenyl)sulfonyl]-2

propenenitrile); BAY 11-7085 ((E)3-[(4-t-Butylphenyl)sulfonyl]-2-propenenitrile); (E)-Capsaicin;

Aurothiomalate (ATM or AuTM); Evodiamine; Hypoestoxide; IKK Inhibitor Ill (BMS-345541); IKK

Inhibitor VII; IKK Inhibitor X; IKK Inhibitor II; IKK-2 Inhibitor IV; IKK-2 Inhibitor V; IKK-2 Inhibitor VI;

IKK-2 Inhibitor (SC-514); IkB Kinase Inhibitor Peptide; IKK-3 Inhibitor IX; ARRY-797 (Array BioPharma);

SB-220025 (5-(2-Amino-4-pyrimidinyl)-4-(4-fluorophenyl)--(4-piperidinlyl)imidazole); SB-239063

(trans-4-[4-(4-Fluorophenyl)-5-(2-methoxy-4-pyrimidinyl) -H-imidazol-1-ylJcyclohexanol); SB-202190 (4-(4-Fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)IH-imidazole); JX-401 (-[2-Methoxy-4

(methylthio)benzoyl]-4-(phenylmethyl)piperidine); PD- 169316 (4-(4-Fuoropheny)-2-(4

nitrophenyl)-5-(4-pyridyl)-IH-imidazole); SKF-86002 (6-(4-Fluorophenyl)-2,3-dihydro-5-(4

pyridinyl)imidazo[2,1-b]thiazole dihydrochloride); SB-200646 (N-(l -Methyl- IH-indo-5-y)-N'-3

pyridinylurea); CMPD-l (2'-Fluoro-N-(4-hydroxyphenyl)-[1,I'-biphenyl]-4-butanamide); EO- 1428 ((2

Methylphenyl)-[4-[(2-amino-4-bromophenyl)amino]-2-ch lorophenyl]methanone) ;SB-253080 (4- [5

(4-Fluorophenyl)-2- [4-(methylsulfonyl)phenyl] - IH-i midazol-4-yl]pyridine); SD-169 (IH-Indole-5

carboxamide); SB-203580 (4-(4-Fluorophenyl)-2-(4-methylsulfinyl phenyl)-5-(4-pyridyl) IH-imidazole); TZP-101 (Tranzyme Pharma); TZP- 102 (Tranzyme Pharma); GHRP-6 (growth hormone-releasing

peptide-6); GHRP-2 (growth hormone-releasing peptide-2); EX-1314 (Elixir Pharmaceuticals); MK-677

(Merck); L-692,429 (Butanamide, 3-amino-3-methyl-N-(2,3,4,5-tetrahydro-2-oxo--((2'-(IH-tetrazo-5

yl)(1,I'-biphenyl)-4-yl)methyl)-IH-1-benzazepin-3-yl)-, (R)-); EP1572 (Aib-DTrp-DgTrp-CHO); diltiazem;

metabolites of diltiazem; BRE (Brain and Reproductive organ-Expressed protein); verapamil;

nimodipine; diltiazem; omega-conotoxin; GVIA; amlodipine; felodipine; lacidipine; mibefradil; NPPB

(5-Nitro-2-(3-phenylpropylamino)benzoic Acid); flunarizine; erythropoietin; pipeline; hemin; brazilin;

z- VAD-FMK (Benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone); z-LEHD-FMK

(benzyloxycarbonyl-Leu-Glu(OMe)-His-Asp(OMe)-fluoromethylketone); B-D-FMK (boc

aspartyl(Ome)-fluoromethylketone); Ac-LEHD-CHO (N-acetyl-Leu-Glu-His-Asp-CHO); Ac-IETD-CHO (N

acetyl-ile-Glu-Thr-Asp-CHO); z-IETD-FMK (benzyloxycarbonyl-Ile-Glu(OMe)-Thr-Asp(OMe)

fluoromethy Iketone); FAM-LEHD-FMK (benzyloxycarbonyl Leu-Glu-His-Asp-fluoromethyl ketone);

FAM-LETD-FMK (benzyloxycarbonyl Leu-Glu-Thr-Asp-fluoromethyl ketone); Q-VD-OPH (Quinoline Val- ASp-CH2-0-Ph); XIAP; clAP-1; cIAP-2; ML-IAP; ILP-2; NAIP; Survivin; Bruce; IAPL-3; fortilin;

leupeptine; PD-150606 (3-(4-lodophenyl)-2-mercapto-(Z)-2-propenoic acid); MDL-28170 (Z-Val-Phe

CHO); calpeptin; acetyl-calpastatin; MG 132 (N-[(phenylmethoxy)carbonyl]-L-leucyl-N-[(IS)--formyl-3

-methylbutyl]-L-leucinamide); MYODUR; BN 82270 (Ipsen); BN 2204 (psen); AHLi-11 (Quark

Pharmaceuticals), an mdm2 protein, pifithrin-a (1-(4-Methylphenyl)-2-(4,5,6,7-tetrahydro-2-imino

3(2H)-benzothiazolyl)ethanone); trans-stilbene; cis-stilbene; resveratrol; piceatannol; rhapontin;

deoxyrhapontin; butein; chalcon; isoliquirtigen; butein; 4,2',4'-trihydroxychalcone; 3,4,2',4',6'

pentahydroxychalcone; flavone; morin; fisetin; luteolin; quercetin; kaempferol; apigenin; gossypetin;

myricetin; 6-hydroxyapigenin; 5-hydroxyflavone; 5,7,3',4',5'-pentahydroxyflavone; 3,7,3',4',5'

pentahydroxyflavone; 3,6,3',4'-tetrahydroxyflavone; 7,3',4',5'-tetrahydroxyflavone; 3,6,2',4'

tetrahydroxyflavone; 7,4'-dihydroxyflavone; 7,8,3',4'-tetrahydroxy flavone; 3,6,2', 3

tetrahydroxyflavone; 4'-hydroxyflavone; 5-hydroxyflavone; 5,4'-dihydroxyflavone; 5,7

dihydroxyflavone; daidzein; genistein; naringenin; flavanone; 3,5,7,3',4'-pentahydroxyflavanone; pelargonidin chloride; cyanidin chloride; delphinidin chloride; (-)-epicatechin (Hydroxy Sites:

3,5,7,3',4A; (-)-catechin (Hydroxy Sites: 3,5,7,3',40; (-)-gallocatechin (Hydroxy Sites: 3,5,7,3 ',4',50

(+)-catechin (Hydroxy Sites: 3,5,7,3',4A; (+)-epicatechin (Hydroxy Sites: 3,5,7,3',41J; Hinokitiol (b

Thujaplicin; 2-hydroxy-4-isopropyl-2,4,6-cycloheptatrien- 1 -one); L-(+)-Ergothioneine ((S)-a-Carboxy

2,3-dihydro-N,N,N-trimethyl-2-thioxo-IH-imidazole4-ethanaminium inner salt); Caffeic Acid Phenyl

Ester; MCI-186 (3-Methyl-l-phenyl-2-pyrazolin-5-one); HBED (N,N'-Di-(2

hydroxybenzyAethylenediamine-HN'-diacetic acid«H20); Ambroxol (trans-4-(2-Amino-3,5

dibromobenzylamino)cyclohexane-HCI; and U-83836E ((-)-2-((4-(2,6-di-1-Pyrrolidinyl-4-pyrimidinyl)-l piperzainyl)methyl)-3,4-dihydro-2,5,7,8-tetramethyl-2H-1-benzopyran-6-ol*2HCI); /5-1 -5-methyl

nicotinamide-2'-deoxyribose; /S-D-1'-5-methyl-nico-tinamide-2'-deoxyribofuranoside; /3-1'-4,5

dimethyl-nicotinamide-2'-de-oxyribose; /3-D-l'-4,5-dimethyl-nicotinamide-2 '-deoxyribofuranoside;

1 -Naphthyl PP 1( 1 -( 1, 1 -Dimethyl ethyl)-3 -( 1 -naphthalenyl)-H-pyrazolo[3, 4-d]pyrimidin-4

amine); Lavendustin A (5-[[(2,5-Dihydroxyphenyl)methyl][(2-hydroxyphenyl)methy I]amino]-2

hydroxybenzoic acid); MNS (3,4-Methylenedioxy-b-nitrostyrene) ; PP 1( 1 -( 1, 1 -Dimethylethyl)- 1

-(4-methylphenyl)- 1 H-pyrazolo[3, 4-d]pyrimidin-4-amine); PP2 (3-(4-chlorophenyl) 1-( 1,1

dimethylethyl)- IH-pyrazolo[3,4-d]pyrimidin-4-amine); KX1-004 (Kinex); KX1-005 (Kinex); KX1-136

(Kinex); KX1-174 (Kinex); KX1-141 (Kinex); KX2-328 (Kinex); KXI-306 (Kinex); KX1-329 (Kinex); KX2-391

(Kinex); KX2-377 (Kinex); ZD4190 (Astra Zeneca; N-(4-bromo-2-fluorophenyl)-6-methoxy-7-(2-(IH

1,2,3-triazol-1-yl)ethoxy)quinazolin-4-amine); AP22408 (Ariad Pharmaceuticals); AP23236 (Ariad

Pharmaceuticals); AP23451 (Ariad Pharmaceuticals); AP23464 (Ariad Pharmaceuticals); AZD0530

(Astra Zeneca); AZM475271 (M475271; Astra Zeneca); Dasatinib (N-(2-chloro-6-methylphneyl)-2-(6 (4-(2-hydroxyethyl)-piperazin-1-yl)-2-methylpyrimidin-4-ylamino) thiazole-5-carboxamide); GN963

(trans-4-(6,7-dimethoxyquinoxalin-2ylamino)cyclohexanol sulfate); Bosutinib (4-((2,4-dichloro-5

methoxyphenyl)amino)-6-methoxy-7-(3-(4-methyl-1-piperazinyl)propoxy)-3-quinolinecarbonitrile); or

combinations thereof.

VII. Fluorescent Moieties

[00286] In some embodiments, the human neuron or nerve targeting molecule further comprises a fluorescent moiety (e.g., a fluorescent protein, peptide, or fluorescent dye molecule). All

fluorescent moieties are encompassed within the term "fluorescent moiety." Specific examples of

fluorescent moieties given herein, are illustrative and are not meant to limit the fluorescent moieties for use with the targeting molecules disclosed herein. In some embodiments, the human neuron or

nerve targeting molecule further comprises a cargo. In some embodiments, the human neuron or

nerve targeting molecule comprises a peptide sequence selected from the group consisting of:

DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC

DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC

(HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20),

QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4;

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), and 5FAM

QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104). In some embodiments the targeting molecule comprises a peptide selected from the group consisting of:

NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2; SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC

(HNP401-N-2; SEQ ID NO:7), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS

(HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21),

and 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104). In some

embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ

ID NO:1). In some embodiments, the targeting molecule comprises the peptide

WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In some embodiments, the targeting molecule

comprises the peptide DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3). In some embodiments, the

targeting molecule comprises the peptide Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4). In some embodiments, the targeting molecule comprises the peptide Ac

WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5). In some embodiments, the

targeting molecule comprises the peptide Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ

ID NO:6). In some embodiments, the targeting molecule comprises the peptide QVPWEEPYYVVKKSS

(HNP401-N-2; SEQ ID NO:20). In some embodiments, the targeting molecule comprises the peptide

QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21). In some embodiments, the

targeting molecule comprises the peptide Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker;

SEQ ID NO:7). In some embodiments, the targeting molecule comprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22). In some embodiments, the targeting molecule

comprises the peptide Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8). In

some embodiments, the targeting molecule comprises the peptide EEPYYVVKKSS (HNP401-N-6; SEQ

ID NO:23). In some embodiments, the targeting molecule comprises the peptide Ac

EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9). In some embodiments, the targeting

molecule comprises the peptide PYYVVKKSS (HNP401-N-8; SEQ ID NO:24). In some embodiments,

the targeting molecule comprises the peptide Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ

ID NO:10). In some embodiments, the targeting molecule comprises the peptide

SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25). In some embodiments, the targeting molecule

comprises the peptide Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11). In

some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYYVV (HNP401-C-4;

SEQ ID NO:26). In some embodiments, the targeting molecule comprises the peptide Ac

SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12). In some embodiments, the targeting molecule comprises the peptide SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27). In some

embodiments, the targeting molecule comprises the peptide Ac-SGQVPWEEPYYGGC (HNP401-C-6

with GGC linker; SEQ ID NO:13). In some embodiments, the targeting molecule comprises the

peptide SGQVPWEEP (HNP401-C-8; SEQ ID NO:28). In some embodiments, the targeting molecule

NO:16). In some embodiments, the targeting molecule comprises the peptide 5FAM

the targeting molecule comprises a peptide that is not Ac-SHSNTQTLAKAPEHTGC (Ac-NP41 with GC

linker; SEQ ID NO:17). In some embodiments, the targeting molecule comprises a peptide that is not

SHSNTQTLAKAPEHTGC (NP41 with GC linker; SEQ ID NO:18). In some embodiments, the peptide is

not NTQTLAKAPEHT (NP41; SEQ ID NO:19).

[00287] In some embodiments, the peptide or aptamer is directly bound to a fluorescent moiety. In some embodiments, the peptide or aptamer is indirectly (e.g., via a linker) bound to a

fluorescent moiety. In some embodiments, the peptide or aptamer is bound to a fluorescent moiety

at its N-terminus, at its C-terminus, or at an internal position (e.g., to an internal amino acid) of the peptide or aptamer. In some embodiments, two or more peptides or aptamers are directly or indirectly bound to a single fluorescent moiety.

[00288] Examples of fluorescent dyes include, but are not limited to, xanthenes (e.g., rhodamines, rhodols and fluoresceins, and their derivatives); bimanes; coumarins and their

derivatives (e.g., umbelliferone and aminomethyl coumarins); aromatic amines (e.g., dansyl;

squarate dyes); benzofurans; fluorescent cyanines; carbazoles; dicyanomethylene pyranes;

polymethine; oxabenzanthrane; xanthene; pyrylium; carbostyl; perylene; acridone; quinacridone;

rubrene; anthracene; coronene; phenanthrecene; pyrene; butadiene; stilbene; porphyrin; pthalocyanine; lanthanide metal chelate complexes; rare-earth metal chelate complexes; and

derivatives of such dyes.

[00289] In some embodiments, the fluorescent moiety is a fluorescein dye. Examples of fluorescein dyes include, but are not limited to, 5-carboxyfluorescein, fluorescein-5-isothiocyanate

and 6-carboxyfluorescein, 5,6-dicarboxyfluorescein, 5-(and 6)-sulfofluorescein, sulfonefluorescein,

succinyl fluorescein, 5-(and 6)-carboxy SNARF-1, carboxyfluorescein sulfonate, carboxyfluorescein

zwitterion, carbxoyfluorescein quaternary ammonium, carboxyfluorescein phosphonate,

carboxyfluorescein GABA, 5'(6')-carboxyfluorescein, carboxyfluorescein-cys-Cy5, and fluorescein

glutathione.

[00290] In some embodiments, the fluorescent moiety is a rhodamine dye. Examples of rhodamine dyes include, but are not limited to, tetramethylrhodamine-6-isothiocyanate, 5

carboxytetramethylrhodamine, 5-carboxy rhodol derivatives, carboxy rhodamine 110, tetramethyl

and tetraethyl rhodamine, diphenyldimethyl and diphenyldiethyl rhodamine, dinaphthyl rhodamine,

rhodamine 101 sulfonyl chloride (sold under the tradename of TEXAS RED).

[00291] In some embodiments, the fluorescent moiety is a cyanine dye. Examples of cyanine dyes include, but are not limited to, Cy3, Cy3B, Cy3.5, Cy5, Cy5.5, Cy 7.

[00292] In some embodiments, the fluorescent moiety is a peptide. In some embodiments, the fluorescent moiety is Green Fluorescent Protein (GFP). In some embodiments, the fluorescent

moiety is a derivative of GFP (e.g., EBFP, EBFP2, Azurite, mKalamal, ECFP, Cerulean, CyPet, YFP, Citrine, Venus, YPet).

[00293] Fluorescent labels are detected by any suitable method. For example, a fluorescent label may be detected by exciting the fluorochrome with the appropriate wavelength of light and detecting the resulting fluorescence, e.g., by microscopy, visual inspection, via photographic film, by the use of electronic detectors such as charge coupled devices (CCDs), photomultipliers, etc.

[00294] In some embodiments, the fluorescent moiety is conjugated to high molecular weight molecule, such as water soluble polymers including, but not limited to,dextran, PEG, serum albumin, or poly(amidoamine) dendrimer.

[00295] Exemplary targeting molecules according to the present invention include: 5FAM QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104), Ac SGQVPWEEPYYVVKKSSGGC-5FAM (HNP401 with GGC linker; SEQ ID NO:105), Ac WEYHYVDLNWTSQHPQGGC-5FAM (HNP402 with GGC linker; SEQ ID NO:106), Ac DLPDIIWDFNWETAGGC-5FAM (HNP403 with GGC linker; SEQ ID NO:107), Ac QVPWEEPYYVVKKSSGGC-5FAM (HNP401-N-2 with GGC linker; SEQ ID NO:108), Ac PWEEPYYVVKKSSGGC-5FAM (HNP401-N-4 with GGC linker; SEQ ID NO:109), Ac-EEPYYVVKKSSGGC 5FAM (HNP401-N-6 with GGC linker; SEQ ID NO:110), Ac-PYYVVKKSSGGC-5FAM (HNP401-N-8 with GGC linker; SEQ ID NO:111), Ac-SGQVPWEEPYYVVKKGGC-5FAM (HNP401-C-2 with GGC linker; SEQ ID NO:112), Ac-SGQVPWEEPYYVVGGC-5FAM (HNP401-C-4 with GGC linker; SEQ ID NO:113), Ac SGQVPWEEPYYGGC-5FAM (HNP401-C-6 with GGC linker; SEQ ID NO:114), and Ac-SGQVPWEEPGGC 5FAM (HNP401-C-8 with GGC linker; SEQ ID NO:115).

Vill. Linkers

[00296] In some embodiments, a cargo (e.g., a fluorescent moiety, photosensitizing agent, or drug) is directly attached to the human neuron or nerve targeting molecule, e.g. at the end of the targeting peptide. Alternatively, in some embodiments, a cargo (e.g., a fluorescent moiety or drug) is indirectly attached to a targeting molecule disclosed herein (e.g., via a linker). In some embodiments, the human neuron or nerve targeting molecule further comprises a cargo. In some embodiments, the human neuron or nerve targeting molecule comprises a peptide sequence selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac DLPDIIWDFNWETAGGC (HNP40 with GGC linker 3; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20),

SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS (HNP401-N-8; SEQ ID

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), and 5FAM

DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), and 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2

with GG linker; SEQ ID NO:104). In some embodiments, the targeting molecule comprises a peptide

selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), Ac

SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), and Ac

QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7). In some embodiments, the

targeting molecule comprises the peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In some

embodiments, the targeting molecule comprises the peptide WEYHYVDLNWTSQHPQ (HNP 402; SEQ

ID NO:2). In some embodiments, the targeting molecule comprises the peptide DLPDIIWDFNWETA

(HNP 403; SEQ ID NO:3). In some embodiments, the targeting molecule comprises the peptide Ac

targeting molecule comprises the peptide Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5). In some embodiments, the targeting molecule comprises the peptide Ac

targeting molecule comprises the peptide Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker;

SEQ ID NO:11). In some embodiments, the targeting molecule comprises the peptide

SEQ ID NO:27). In some embodiments, the targeting molecule comprises the peptide Ac SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13). In some embodiments, the

embodiments, the targeting molecule comprises the peptide Ac-SGQVPWEEPGGC (HNP401-C-8 with

GGC linker; SEQ ID NO:14). In some embodiments, the targeting molecule comprises the peptide

NO:104). In some embodiments, the targeting molecule comprises a peptide that is not Ac

SHSNTQTLAKAPEHTGC (Ac-NP41 with GC linker; SEQ ID NO:17). In some embodiments, the targeting

molecule comprises a peptide that is not SHSNTQTLAKAPEHTGC (NP41 with GC linker; SEQ ID

NO:18). In some embodiments, the targeting molecule comprises a peptide that is not

NTQTLAKAPEHT (SEQ ID NO:19).

[00297] As used herein, a "linker" is any molecule capable of binding (e.g., covalently) to a targeting molecule disclosed herein. Linkers include, but are not limited to, straight or branched

chain carbon linkers, heterocyclic carbon linkers, amino acid linkers (e.g., D- or L-amino acid),

lipophilic residues, peptide linkers, peptide nucleic acid linkers, hydrazone linkers, SPDB disulfide,

sulfo-SPDB, maleimidomethyl cyclohexane-1-carboxylate (MCC), aminohexanoic acid linkers, and

polyether linkers (e.g., PEG). For example, poly(ethylene glycol) linkers are available from Quanta

Biodesign, Powell, OH. These linkers optionally have amide linkages, sulfhydryl linkages, or hetero

functional linkages.

[00298] In some embodiments, the linker binds to a targeting molecule disclosed herein by a covalent linkage. In some embodiments, the covalent linkage comprises an ether bond, thioether

bond, amine bond, amide bond, carbon-carbon bond, carbon-nitrogen bond, carbon-oxygen bond,

or carbon-sulfur bond.

[00299] In some embodiments, the linker is flexible. In some embodiments, the linker is rigid.

[00300] In some embodiments, the linker comprises a linear structure. In some embodiments, the linker comprises a non-linear structure. In some embodiments, the linker

comprises a branched structure. In some embodiments, the linker comprises a cyclic structure.

[00301] In some embodiments, the linker is an alkyl. In some embodiments, the linker is heteroalkyl.

[00302] In some embodiments, the linker is an alkylene. In some embodiments, the linker is an alkenylene. In some embodiments, the linker is an alkynylene. In some embodiments, the linker is

a heteroalkylene.

[00303] An "alkyl" group refers to an aliphatic hydrocarbon group. The alkyl moiety may be a saturated alkyl or an unsaturated alkyl. Depending on the structure, an alkyl group can be a

monoradical or a diradical (i.e., an alkylene group).

[00304] The"alkyl" moiety may have 1to 10 carbon atoms (whenever it appears herein, a numerical range such as "1 to 10" refers to each integer in the given range; e.g., "1 to 10 carbon

atoms" means that the alkyl group may consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms,

etc., up to and including 10 carbon atoms, although the present definition also covers the

occurrence of the term "alkyl" where no numerical range is designated). The alkyl group could also be a "lower alkyl" having 1 to 6 carbon atoms. The alkyl group of the compounds described herein

may be designated as "Ci-C4 alkyl" or similar designations. By way of example only, "C1-C4 alkyl"

indicates that there are one to four carbon atoms in the alkyl chain, i.e., the alkyl chain is selected

from methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec -butyl, and t-butyl. Typical alkyl groups include, but are in no way limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl, pentyl, hexyl, ethenyl, propenyl, butenyl, and the like.

[00305] In some embodiments, the linker comprises a ring structure (e.g., an aryl). As used herein, the term "ring" refers to any covalently closed structure. Rings include, for example,

carbocycles (e.g., aryls and cycloalkyls), heterocycles (e.g., heteroaryls and non-aromatic

heterocycles), aromatics (e.g. aryls and heteroaryls), and non-aromatics (e.g., cycloalkyls and non

aromatic heterocycles). Rings can be optionally substituted. Rings can be monocyclic or polycyclic.

[00306] As used herein, the term "aryl" refers to an aromatic ring wherein each of the atoms forming the ring is a carbon atom. Aryl rings can be formed by five, six, seven, eight, nine, or more

than nine carbon atoms. Aryl groups can be optionally substituted. Examples of aryl groups include,

but are not limited to phenyl, naphthalenyl, phenanthrenyl, anthracenyl, fluorenyl, and indenyl.

Depending on the structure, an aryl group can be a monoradical or a diradical (i.e., an arylene

group).

[00307] The term "cycloalkyl" refers to a monocyclic or polycyclic non-aromatic radical, wherein each of the atoms forming the ring (i.e. skeletal atoms) is a carbon atom. Cycloalkyls may be

saturated, or partially unsaturated. Cycloalkyl groups include groups having from 3 to 10 ring atoms.

Cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[00308] In some embodiments, the ring is a cycloalkane. In some embodiments, the ring is a cycloalkene.

[00309] In some embodiments, the ring is an aromatic ring. The term "aromatic" refers to a planar ring having a delocalized n-electron system containing 4n+2 1 electrons, where n is an

integer. Aromatic rings can be formed from five, six, seven, eight, nine, or more than nine atoms.

Aromatics can be optionally substituted. The term "aromatic" includes both carbocyclic aryl (e.g.,

phenyl) and heterocyclic aryl (or "heteroaryl" or "heteroaromatic") groups (e.g., pyridine). The term

includes monocyclic or fused-ring polycyclic (i.e., rings which share adjacent pairs of carbon atoms)

groups.

[00310] In some embodiments, the ring is a heterocycle. The term "heterocycle" refers to heteroaromatic and heteroalicyclic groups containing one to four heteroatoms each selected from

0, S and N, wherein each heterocyclic group has from 4 to 10 atoms in its ring system, and with the

proviso that the ring of said group does not contain two adjacent 0 or S atoms. Non-aromatic

heterocyclic groups include groups having only 3 atoms in their ring system, but aromatic heterocyclic groups must have at least 5 atoms in their ring system. The heterocyclic groups include

benzo-fused ring systems. An example of a 3-membered heterocyclic group is aziridinyl. An example

of a 4-membered heterocyclic group is azetidinyl (derived from azetidine). An example of a 5

membered heterocyclic group is thiazolyl. An example of a 6-membered heterocyclic group is

pyridyl, and an example of a 10-membered heterocyclic group is quinolinyl. Examples of non

aromatic heterocyclic groups are pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl,

tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino,

thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl,

oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl,

2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl,

dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl,

3-azabicyclo[4.1.0]heptanyl, 3H-indolyl and quinolizinyl. Examples of aromatic heterocyclic groups

are pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl,

benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl,

pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl,

benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and faropyridinyl. The

foregoing groups, may be C-attached or N-attached where such is possible. For instance, a group

derived from pyrrole may be pyrrol- 1-yl (N-attached) or pyrrol-3-yl (C-attached). Further, a group

derived from imidazole may be imidazol-1-yl or imidazol-3-yl (both N-attached) or imidazol-2-yl,

imidazol-4-yl or imidazol-5-yl (all C-attached). The heterocyclic groups include benzo-fused ring

systems and ring systems substituted with one or two oxo (=0) moieties such as pyrrolidin-2-one.

Depending on the structure, a heterocycle group can be a monoradical or a diradical (i.e., a

heterocyclene group).

[00311] In some embodiments, the ring is fused. The term "fused" refers to structures in which two or more rings share one or more bonds, hi some embodiments, the ring is a dimer. In

some embodiments, the ring is a trimer. In some embodiments, the ring is a substituted.

[00312] The term "carbocyclic" or "carbocycle" refers to a ring wherein each of the atoms forming the ring is a carbon atom. Carbocycle includes aryl and cycloalkyl. The term thus distinguishes carbocycle from heterocycle ("heterocyclic") in which the ring backbone contains at least one atom which is different from carbon (i.e., a heteroatom). Heterocycle includes heteroaryl and heterocycloalkyl. Carbocycles and heterocycles can be optionally substituted.

[00313] In some embodiments, the linker is substituted. The term "optionally substituted" or "substituted" means that the referenced group may be substituted with one or more additional

group(s) individually and independently selected from Ci-Cealkyl, C3-Cgcycloalkyl, aryl, heteroaryl,

C2-C6heteroalicyclic, hydroxy, Ci-C6alkoxy, aryloxy, Ci-C6alkylthio, arylthio, Ci-C6alkylsulfoxide,

arylsulfoxide, Ci-C6alkylsulfone, arylsulfone, cyano, halo, C2-C8acyl, C2-C8acyloxy, nitro, Ci C6haloalkyl, Ci-C6fluoroalkyl, and amino, including Ci-C6alkylamino, and the protected derivatives

thereof. By way of example, an optional substituents may be LSRS, wherein each Ls is independently

selected from a bond, -0-,-C(=)-, -S-, -S(=)-, -S(0)2-, -NH-, -NHC(O)-, -C(O)NH-, S(O)2NH-,

NHS()2-, -OC(O)NH-, -NHC(O)O-, -(CpC6alkyl)-, or -(C2-C6alkenyl)-; and each Rs is independently

selected from H, (Ci-C4alkyl), (C3-C8cycloalkyl), heteroaryl, aryl, and Ci-C6heteroalkyl. Optionally

substituted non-aromatic groups may be substituted with one or more oxo (=0). The protecting

groups that may form the protective derivatives of the above substituents are known to those of skill

in the art.

[00314] In some embodiments, a bifunctional linker having one functional group reactive with a group on one molecule (e.g., a targeting molecule), and another group reactive on the other

molecule (e.g., a fluorescent moiety or a drug), is used to form the desired conjugate. Alternatively,

in some embodiments, derivatization is performed to provide functional groups. Thus, for example,

procedures for the generation of free sulfhydryl groups on peptides are also known (See U.S. Pat.

No. 4,659,839). A linker may alternatively comprise a heterobifunctional crosslinker comprising two

or more different reactive groups that form a heterocyclic ring that can interact with a targeting

molecule. For example, a heterobifunctional crosslinker such as cysteine may comprise an amine

reactive group and a thiol-reactive group can interact with an aldehyde on a derivatized targeting

molecule. Additional combinations of reactive groups suitable for heterobifunctional crosslinkers

include, for example, amine- and sulfhydryl reactive groups; carbonyl and sulfhydryl reactive groups;

amine and photoreactive groups; sulfhydryl and photoreactive groups; carbonyl and photoreactive

groups; carboxylate and photoreactive groups; and arginine and photoreactive groups. Examples of

a heterobifunctional crosslinker include N-Succinimidyl 4-(2-pyridyldithio)butanoate (SPDB) and

maleimidomethyl cyclohexane-1-carboxylate (MCC).

[00315] In some embodiments, a peptide linker consisting of one or more amino acids is used to join the targeting molecule and a fluorescent moiety or drug. Generally the peptide linker

will have no specific biological activity other than to join the molecules or to preserve some

minimum distance or other spatial relationship between them. However, the constituent amino

acids of the linker may be selected to influence some property of the molecule such as the folding,

net charge, or hydrophobicity. In some embodiments the peptide linker is relatively short, typically

less than about 10 amino acids, preferably less than about 8 amino acids and more preferably less

than 5 amino acids. Non-limiting illustrative examples include glycine and glycine-serine linkers

which can be added to the C-terminus of a targeting peptide. In some embodiments, a peptide

linker is a glycine-glycine-glycine-cysteine (GGGC) linker, a glycine-glycine-cysteine (GGC) linker, a

glycine-glycine (GG) linker, or a cysteine (C) linker. In some embodiments, the GGGC, GGC, GG, or C

linker is added to the C-terminus of a targeting peptide.

IX. Further Modifications

[00316] In some embodiments, the human neuron or nerve targeting molecules of the present invention are optionally conjugated to high molecular weight molecules that increase the

multivalency and avidity of labeling. In some embodiments, the high molecular weight molecules are

water-soluble polymers. Examples of suitable water-soluble polymers include, but are not limited to,

peptides, saccharides, poly(vinyls), poly(ethers), poly(amines), poly(carboxylic acids) and the like. In

some embodiments, the water-soluble polymers isdextran, polyethylene glycol (PEG),

polyoxyalkylene, polysialic acid, starch, or hydroxyethyl starch. Any suitable method is used to

conjugate peptides to water-soluble polymers (see, Hermanson G., Bioconjugate Techniques 2nd

Ed., Academic Press, Inc. 2008). In some embodiments, the human neuron or nerve targeting

molecule further comprises a cargo. In some embodiments, the human neuron or nerve targeting

molecule comprises a peptide sequence selected from the group consisting of:

NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac

SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),

QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22), EEPYYVVKKSS (HNP401-N-6;

SEQ ID NO:23), PYYVVKKSS (HNP401-N-8; SEQ ID NO:24), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID

SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),

ID NO:104). In some embodiments the targeting molecule comprises a peptide selected from the

group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP

402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), and 5FAM-QVPWEEPYYVVKKSSGG

NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104). In some embodiments, the targeting molecule comprises a peptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID

NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac

C-2; SEQ ID NO:25), Ac-QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), and Ac

DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6). In some embodiments, the targeting molecule comprises the peptide QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some

targeting molecule comprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22). In some embodiments, the targeting molecule comprises the peptide Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8). In some embodiments, the targeting molecule comprises the peptide

EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23). In some embodiments, the targeting molecule comprises the peptide Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9). In some

targeting molecule comprises the peptide SGQVPWEEP (HNP401-C-8; SEQ ID NO:28). In some embodiments, the targeting molecule comprises the peptide Ac-SGQVPWEEPGGC (HNP401-C-8 with

NTQTLAKAPEHT (SEQ ID NO:19).

[00317] In some embodiments, the targeting molecules of the present invention are conjugated to factors having neurotrophic properties (e.g., neurotrophic proteins such as nerve

growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3),

neurotrophin-4 (NT-4), glial cell line-derived neurotrophic factor (GDNF), ciliary neurotrophic factor

(CNTF) as well as non-protein small molecules with neurotrophic properties).

[00318] In some embodiments, the targeting molecules of the present invention are modified to increase solubility. Peptide modifications that increase solubility include addition of hyphilic amino acid(s), a PEG moiety, or both. In some embodiments, a PEG moiety is 8-Amino-3,6 dioxaoctanoic acid (AEEA); 12-amino-4,7,10-trioxadodecanoic acid; or 15-amino-4,7,10,13 tetraoxapenta-decanoic acid. In some embodiments, about one to ten (e.g., one, two, three, four, five, six, seven, eight, nine, or ten) hydrophilic amino acids may be added to the N-terminus, C terminus, an internal position, or any combination thereof, of the targeting molecule to increase solubility. Hydrophilic amino acids include D, E, H, K, N, Q, R, S, T, and G. In some embodiments, the targeting molecule comprises a K, KK, G, or GG at the N-terminus or C-terminus.

X. Multidomain Targeting Molecules

[00319] In certain embodiments, the human neuron or nerve targeting molecules provided herein are multidomain neuron or nerve targeting molecules comprising two or more neuron or

nerve targeting peptides, wherein the first peptide comprises SGQVPWEEPYYVVKKSS (HNP 401; SEQ

ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with

GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5),

Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20);

NO:26), SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), or 5FAM

the first peptide comprises SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ

(HNP 402; SEQ ID NO:2), or DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3). In some embodiments, the

first peptide comprises SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC

(HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS

(HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21), or

5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104). In some

embodiments, the first peptide comprises SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In some

embodiments, the first peptide comprises WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In some

embodiments, the first peptide comprises DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3). In some

embodiments, the first peptide comprises Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker;

SEQ ID NO:4). In some embodiments, the first peptide comprises Ac-WEYHYVDLNWTSQHPQGGC

(HNP402 with GGC linker; SEQ ID NO:5). In some embodiments, the first peptide comprises Ac

DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6). In some embodiments, the first

peptide comprises QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some embodiments, the

first peptide comprises QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21). In some

embodiments, the first peptide comprises Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7). In some embodiments, the first peptide comprises PWEEPYYVVKKSS (HNP401-N-4;

SEQ ID NO:22). In some embodiments, the first peptide comprises Ac-PWEEPYYVVKKSSGGC

(HNP401-N-4 with GGC linker; SEQ ID NO:8). In some embodiments, the first peptide comprises

EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23). In some embodiments, the first peptide comprises Ac

EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9). In some embodiments, the first

peptide comprises PYYVVKKSS (HNP401-N-8; SEQ ID NO:24). In some embodiments, the first peptide

comprises Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10). In some embodiments,

the first peptide comprises SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25). In some

embodiments, the first peptide comprises Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC

linker; SEQ ID NO:11). In some embodiments, the first peptide comprises SGQVPWEEPYYVV

(HNP401-C-4; SEQ ID NO:26). In some embodiments, the first peptide comprises Ac

SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12). In some embodiments, the first

peptide comprises SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27). In some embodiments, the first peptide comprises Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13). In some

embodiments, the first peptide comprises SGQVPWEEP (HNP401-C-8; SEQ ID NO:28). In some

embodiments, the first peptide comprises Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID

NO:14). In some embodiments, the first peptide comprises DTHAHAKPRVPAFKSV (HNP 404; SEQ ID

NO:16). In some embodiments, the first peptide comprises 5FAM-QVPWEEPYYVVKKSSGG-NH2

(HNP401-N-2 with GG linker; SEQ ID NO:104).

[00320] The two or more neuron or nerve targeting peptides within a multidomain targeting molecule can be the same neuron or nerve targeting peptide, or are preferably different neuron or

nerve targeting peptides. In some embodiments, multidomain targeting molecules comprise a

second peptide comprising: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ

(HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP

404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC

SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac

NO:11), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20);

PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120),

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), 5FAM

QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104), SHSSEFPRSWDMETN

(HNP301; SEQ ID NO:29); SHSMLPSVLD (HNP303; SEQ ID NO:30); SHSTMKTLSL (HNP305; SEQ ID

NO:31); VAPTKAPLHSPS (NP121; SEQ ID NO:32), NNLKTGTSAPTG (NP122; SEQ ID NO:33),

HKTAQWPFIAFR (NP123; SEQ ID NO:34), RLTNAPAYQAPA (NP124; SEQ ID NO:35), MQNPLNGKPGR

(NP125; SEQ ID NO:36), THYSRSLTDGTR (NP126; SEQ ID NO:37), YPSPNRPPNLTN (NP127; SEQ ID

NO:38), or NTQTLAKAPEHTG (NP117; SEQ ID NO:39).

[00321] In some emboidments, the first neuron or nerve targeting peptide is selected from the group consisting of: QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG

(HNP401-N-2 with GG linker; SEQ ID NO:21), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), and

5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104). In some

embodiments, the second peptide is selected from the group consisting of: SGQVPWEEPYYVVKKSS

(HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP

403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC

(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker;

SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac

SEQ ID NO:9), Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac

SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),

QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6;

SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),

NO:104), SHSSEFPRSWDMETN (HNP301; SEQ ID NO:29); SHSMLPSVLD (HNP303; SEQ ID NO:30);

SHSTMKTLSL (HNP305; SEQ ID NO:31); VAPTKAPLHSPS (NP121; SEQ ID NO:32), NNLKTGTSAPTG

(NP122; SEQ ID NO:33), HKTAQWPFIAFR (NP123; SEQ ID NO:34), RLTNAPAYQAPA (NP124; SEQ ID

NO:35), MQNPLNGKPGR (NP125; SEQ ID NO:36), THYSRSLTDGTR (NP126; SEQ ID NO:37), YPSPNRPPNLTN (NP127; SEQ ID NO:38), or NTQTLAKAPEHTG (NP117; SEQ ID NO:39).

[00322] In some embodiments, the first neuron or nerve targeting peptide comprises QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some embodiments, the first neuron or nerve

targeting peptide is QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20).In some embodiments, the second peptide is selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID

DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20);

NO:26), SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), and SGQVPWEEP (HNP401-C-8; SEQ ID NO:28),

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), SHSSEFPRSWDMETN

NO:31); VAPTKAPLHSPS (NP121; SEQ ID NO:32), NNLKTGTSAPTG (NP122; SEQ ID NO:33),

NO:38), or NTQTLAKAPEHTG (NP117; SEQ ID NO:39).

[00323] In some embodiments, the first neuron or nerve targeting peptide comprises QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21). In some embodiments, the first

neuron or nerve targeting peptide is QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID

NO:21). In some embodiments, the second peptide is selected from the group consisting of:

NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac

6 with GGC linker; SEQ ID NO:9), Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-SGQVPWEEPYYVVGGC

SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),

SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),

GG linker; SEQ ID NO:124), 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104), SHSSEFPRSWDMETN (HNP301; SEQ ID NO:29); SHSMLPSVLD (HNP303; SEQ ID NO:30);

NO:35), MQNPLNGKPGR (NP125; SEQ ID NO:36), THYSRSLTDGTR (NP126; SEQ ID NO:37),

YPSPNRPPNLTN (NP127; SEQ ID NO:38), or NTQTLAKAPEHTG (NP117; SEQ ID NO:39).

[00324] In some embodiments, the first neuron or nerve targeting peptide comprises SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25). In some embodiments, the first neuron or nerve

targeting peptide is SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25). In some embodiments, the

second peptide is selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID

DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20);

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124),5FAM

NO:31); VAPTKAPLHSPS (NP121; SEQ ID NO:32), NNLKTGTSAPTG (NP122; SEQ ID NO:33),

(NP125; SEQ ID NO:36), THYSRSLTDGTR (NP126; SEQ ID NO:37), YPSPNRPPNLTN (NP127; SEQ ID NO:38), or NTQTLAKAPEHTG (NP117; SEQ ID NO:39).

[00325] In some embodiments, the first neuron or nerve targeting peptide comprises 5FAM QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104). In some embodiments,

the first neuron or nerve targeting peptide is 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with

GG linker; SEQ ID NO:104). In some embodiments, the second peptide is selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ

NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac

WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC

SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac

NO:11), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20);

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), 5FAM

QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104), SHSSEFPRSWDMETN (HNP301; SEQ ID NO:29); SHSMLPSVLD (HNP303; SEQ ID NO:30); SHSTMKTLSL (HNP305; SEQ ID

NO:31); VAPTKAPLHSPS (NP121; SEQ ID NO:32), NNLKTGTSAPTG (NP122; SEQ ID NO:33),

NO:38), or NTQTLAKAPEHTG (NP117; SEQ ID NO:39).

[00326] In some embodiments, the neuron or nerve targeting peptides within a multidomain targeting molecule are directly bound to each other. In some embodiments, the neuron or nerve

targeting peptides within a multidomain targeting molecule are indirectly bound to each other, e.g.,

via a linker or cargo. In some embodiments, the targeting peptides are arranged in a linear fashion.

In some embodiments, the targeting peptides of a multidomain targeting molecule are arranged in a

branched strucutre. In some embodiments, a multidomain targeting molecule comprises two, three,

four, five, or more neuron or nerve targeting peptides.

XI. Methods of Labeling

[00327] Disclosed herein, in certain embodiments, are methods of labeling a neuron or nerve (or component of either) by contacting a neuron or nerve with a human neuron or nerve targeting

molecule described herein. In some embodiments, the human neuron or nerve targeting molecule

further comprises a cargo. In some embodiments, the human neuron or nerve targeting molecule

comprises a peptide sequence selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP

SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC

(HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac

SEQ ID NO:9), Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac

SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker;

SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14),

QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG

linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22), EEPYYVVKKSS (HNP401-N-6;

SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122),

ID NO:104). In some embodiments the targeting molecule comprises a peptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP

402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3). In some embodiments, the

targeting molecule comprises a peptide selected from the group consisting of SGQVPWEEPYYVVKKSS

(HNP 401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11),

Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), SGQVPWEEPYYVVKK

(HNP401-C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), and

NTQTLAKAPEHT (SEQ ID NO:19).

[00328] In some embodiments, a first human neuron or nerve targeting molecule is administered in combination (simultaneously, concurrently, or serially) with a second human neuron

or nerve targeting molecule. In further embodiments, the first targeting molecule, the second

targeting molecule, or both comprise a cargo. In yet further embodiments, the cargo of the first

targeting molecule, the cargo of the second targeting molecule, or both are fluorescent moieities,

which may be the same fluorescent moeities or different fluorescent moieties.

[00329] In some embodiments, a human neuron or nerve targeting molecule is adminstered in combination (simultaneously, concurrently, or serially) with a fluorescent moiety (e.g., fluorescent moiety is not conjugated to the targeting molecule, "free" fluorescent moiety). In some embodiments, the fluorescent moiety is a fluorescein, e.g., carboxyfluorescein.

[00330] In some embodiments, the contacting occurs in vivo. In some embodiments, the contacting occurs in vitro.

[00331] In some embodiments, a neuron or nerve (or component thereof) is labeled for identification during surgery. In some embodiments the surgery is cancer surgery. In some embodiments the cancer is selected from the group consisting of prostate cancer, liver cancer (HCC), colorectal cancer, ovarian cancer, endometrial cancer, breast cancer, pancreatic cancer, stomach cancer, cervical cancer, head and neck cancer, thyroid cancer, testis cancer, urothelial cancer, lung cancer, melanoma, testicular germ cell tumors, mesothelioma, and esophageal cancer. Insome embodiments, the cancer is prostate cancer. In some embodiments, the method comprises administering a targeting molecule disclosed herein to a subject that will undergo surgery. In some embodiments, the method comprises administering a targeting molecule disclosed herein to a subject that is undergoing surgery. In some embodiments, a targeting molecule disclosed herein is administered to a patient systemically. In some embodiments, a targeting molecule disclosed herein is administered to a patient locally.

XII. Drug Delivery

[00332] Disclosed herein, in certain embodiments, are methods of targeted drug delivery. In some embodiments, a human neuron or nerve targeting molecule disclosed herein delivers a drug to a specific target. In some embodiments, a targeting molecule disclosed herein delivers a drug to a neuron or nerve. In some embodiments, the human neuron or nerve targeting molecule further comprises a cargo. In some embodiments, the human neuron or nerve targeting molecule comprises a peptide sequence selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac-WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9), Ac-PYYVVKKSSGGC

SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20),

SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28),

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), and 5FAM

SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), and DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3). In some embodiments, the targeting molecule

NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac

C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20), and QVPWEEPYYVVKKSSGG

(HNP401-N-2 with GG linker; SEQ ID NO:21). In some embodiments, the targeting molecule

comprises the peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In some embodiments, the

targeting molecule comprises the peptide WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In some

NO:3). In some embodiments, the targeting molecule comprises the peptide Ac

SEQ ID NO:5). In some embodiments, the targeting molecule comprises the peptide Ac DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6). In some embodiments, the

Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7). In some embodiments, the targeting molecule comprises the peptide PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22). In some embodiments, the targeting molecule comprises the peptide Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8). In some embodiments, the targeting molecule comprises the peptide EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23). In some embodiments, the targeting molecule comprises the peptide Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9). In some embodiments, the targeting molecule comprises the peptide PYYVVKKSS (HNP401-N-8; SEQ ID

SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13). In some embodiments, the targeting molecule comprises the peptide SGQVPWEEP (HNP401-C-8; SEQ ID NO:28). In some

NTQTLAKAPEHT (SEQ ID NO:19).

[00333] In some embodiments, the drug is an agent that reduces pain (either the perception of pain or activity of a painful stimulant). In some embodiments, the drug is an anesthetic. In some

embodiments, the drug is benzocaine; carticaine; cinchocaine; cyclomethycaine; lidocaine;

prilocaine; propxycaine; proparacaine; tetracaine; tocainide; and trimecaine; or a combination

thereof.

[00334] In some embodiments, the drug is an agent that modulates death (e.g., via apoptosis or necrosis) of a neuron or nerve. In some embodiments, the drug is a cytotoxic agent. In some embodiments, the drug is methotrexate (RHEUMATREX©, Amethopterin); cyclophosphamide (CYTOXAN©);thalidomide (THALIDOMID*); paclitaxel; pemetrexed; pentostatin; pipobroman; pixantrone; plicamycin; procarbazine; proteasome inhibitors (e.g.; bortezomib); raltitrexed; rebeccamycin; rubitecan; SN-38; salinosporamide A; satraplatin; streptozotocin; swainsonine; tariquidar; taxane; tegafur-uracil; temozolomide; testolactone; tbioTEPA; tioguanine; topotecan; trabectedin; tretinoin; triplatin tetranitrate; tris(2-chloroethyl)amine; troxacitabine; uracil mustard; valrubicin; vinblastine; vincristine; vinorelbine; vorinostat; zosuquidar; or a combination thereof. In some embodiments, the drug is a pro-apoptotic agent. In some embodiments, the drug is an anti apoptotic agent. In some embodiments, the drug is selected from minocycline; SB-203580 (4-(4 Fluorophenyl)-2-(4-methylsulfmyl phenyl)-5-(4-pyridyl) IH-imidazole); PD 169316 (4-(4 Fluorophenyl)-2-(4-nitrophenyl)-5-(4-pyridyl)-IH-imidazole); SB 202190 (4-(4-Fluoropheny)-2-(4 hydroxyphenyl)-5-(4-pyridyl)IH-imidazole); RWJ 67657 (4-[4-(4-fluorophenyl)--(3-phenylpropyl)-5-(4 pyridinyl)-IH-imidazol -2-yl]-3-butyn-1-ol); SB 220025 (5-(2-Amino-4-pyrimidinyl)-4-(4-fluorophenyl) 1 -(4-piperidinlyl)imidazole) ; D-JNKI- 1 ((D)-hJIP 175_i 57-DPrO-DPrO-(D)-HIV-TAT57-48); AM-111 (Auris); SP600125 (anthra[1,9-cd]pyrazol-6(2H)-one); JNK Inhibitor I ((L)-HIV-T AT48-57-PP-JBD20); JNK Inhibitor Ill ((L)-HIV-TAT47-57-gaba-c-Jun533-57);AS601245(1,3-benzothiazol-2-y (2-[[2-(3 pyridinyl) ethyl] amino]-4 pyrimidinyl) acetonitrile); JNK Inhibitor VI (H2N-RPKRPTTLNLF-NH2); JNK Inhibitor Vill (N-(4-Amino-5-cyano-6-ethoxypyridin-2-yl)-2-(2,5-dimethoxyphenyl)acetamide); JNK Inhibitor IX(N-(3-Cyano-4,5,6,7-tetrahydro-l-benzothien-2-yl)-l-naphthamide); dicumarol (3,3 ' Methylenebis(4-hydroxycoumarin)); SC-236 (4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-H-pyrazol-l yl]benzene-sulfonamide); CEP-1347 (Cephalon); CEP-11004 (Cephalon); an artificial protein comprising at least a portion of a Bc-2 polypeptide; a recombinant FNK; V5 (also known as Bax inhibitor peptide V5); Bax channel blocker (()--(3,6-Dibromocarbazol-9-yl)-3-piperazin-1-yl-propan 2-ol); Bax inhibiting peptide P5 (also known as Bax inhibitor peptideP5); Kp7-6; FAIM(S) (Fas apoptosis inhibitory molecule-short) ; FAIM(L) (Fas apoptosis inhibitory molecule-long) ; Fas : Fc; FAP- 1; NOK2 ; F2051; FI 926; F2928; ZB4; Fas M3 mAb; EGF; 740 Y-P; SC 3036 (KKHTDDGYMPMSPGVA); PI 3-kinase Activator (Santa Cruz Biotechnology, Inc.); Pam3Cys ((S)-(2,3 bis(palmitoyloxy)-(2RS)-propyl)-N-palmitoyl-(R)-Cys-(S)-Ser(S)-Lys4-OH, trihydrochloride); Actl (NF-kB activator 1); an anti-DcB antibody; Acetyl- 11-keto-b-Boswellic Acid; Andrographolide; Caffeic Acid Phenethyl Ester (CAPE); Gliotoxin; Isohelenin; NEMO-Binding Domain Binding Peptide (DRQIKIWFQNRRMKWKKTALDWSWLQTE); NF-kB Activation Inhibitor (6-Amino-4-(4 phenoxyphenylethylamino)quinazoline); NF-kB Activation Inhibitor11 (4-Methyl-NI-(3 phenylpropyl)benzene-1,2-diamine); NF-kB Activation Inhibitor Ill (3-Chloro-4-nitro-N-(5-nitro-2 thiazolyl)-benzamide); NF-kB Activation hihibitor IV ((E)-2-Fluoro-4'-methoxystilbene); NF-kB

Activation Inhibitor V (5-Hydroxy-(2,6-diisopropylphenyl)-lH-isoindole-1,3-dione); NF-kB SN50 (AA VALLP A VLLALLAP VQRKRQKLMP); Oridonin; Parthenolide; PPM-18 (2-Benzoylamino-1,4

naphthoquinone); Rol06-9920; Sulfasalazine; TIRAP Inhibitor Peptide (RQIKWFNRRMKWKKLQLRD

AAPGG AIVS); Withaferin A; Wogonin; BAY 11-7082 ((E)3-[(4-Methylphenyl)sulfonyl]-2

IKK-2 Inhibitor (SC-514); IkB Kinase Inhibitor Peptide; IKK-3 Inhibitor LX; ARRY-797 (Array

BioPharma); SB-220025 (5-(2-Amino-4-pyrimidinyl)-4-(4-fluorophenyl)- 1 -(4-piperidinlyl)imidazole);

SB-239063 (trans-4-[4-(4-Fluorophenyl)-5-(2-methoxy-4-pyrimidinyl) -H-imidazol-1-yl]cyclohexanol);

SB-202190 (4-(4-Fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl) 1 H-imidazole); JX-401 (-[2

Methoxy-4-(methylthio)benzoyl]-4-(phenylmethyl)piperidine); PD-169316 (4-(4-Fluorophenyl)-2-(4

pyridinyl)imidazo[2,1-b]thiazole dihydrochloride); SB-200646 (N-(I-Methyl-H-indol-5-yl)-N'-3 pyridinylurea); CMPD-I (2'-Fluoro-N-(4-hydroxyphenyl)-[ 1, 1'-biphenyl]-4-butanamide); EO- 1428

((2-Methylphenyl)-[4-[(2-amino-4-bromophenyl)amino]-2-ch lorophenyl]methanone);SB-253080 (4

[5-(4-Fluorophenyl)-2-[4-(methylsulfonyl)phenyl]-1H-i midazol-4-yl]pyridine); SD-169 (IH-Indole-5

carboxamide); SB-203580 (4-(4- Fluorophenyl)-2-(4-methylsulfmyl phenyl)-5-(4-pyridyl) 1 H

imidazole); TZP-101 (Tranzyme Pharma); TZP-102 (Tranzyme Pharma); GHRP-6 (growth hormone

releasing peptide-6); GHRP-2 (growth hormone-releasing peptide-2); EX-1314 (Elixir

Pharmaceuticals); MK-677 (Merck); L-692,429 (Butanamide, 3-amino-3-methyl-N-(2,3,4,5

tetrahydro-2-oxo--((2'-(IH-tetrazo-5-yl)(1,I'-biphenyl)-4-yl)methyl)-lH-1-benzazepin-3-yl)-, (R)-);

EP1572 (Aib-DTrp-DgTc-CHO); diltiazem; metabolites of diltiazem; BRE (Brain and Reproductive

organ-Expressed protein); verapamil; nimodipine; diltiazem; omega-conotoxin; GVIA; amlodipine;

felodipine; lacidipine; mibefradil; NPPB (5-Nitro-2-(3-phenylpropylamino)benzoic Acid); flunarizine;

erythropoietin; piperine; hemin; brazilin; z- V AD-FMK (Benzyloxycarbonyl-Val-Ala-Asp(OMe)

fluoromethylketone); z-LEHD-FMK (benzyloxycarbonyl-Leu-Glu(OMe)- His-Asp(OMe) fluoromethylketone); B-D-FMK (boc-aspartyl(Ome)-fluoromethylketone); Ac-LEHD-CHO (N-acetyl

Leu-Glu-His-Asp-CHO); Ac-IETD-CHO (N-acetyl-ile-Glu-Thr-Asp-CHO); z-IETD-FMK (benzyloxycarbonyl

lle-Glu(OMe)-Thr-Asp(OMe)-fluoromethy Iketone); FAM-LEHD-FMK (benzyloxycarbonyl Leu-Glu-His

Asp-fluoromethyl ketone); FAM-LETD-FMK (benzyloxycarbonyl Leu-Glu-Thr-Asp-fluoromethyl

ketone); Q-VD-OPH (Quinoline- Val- ASp-CH2-0-Ph); XIAP; clAP-1; cAP-2; ML-IAP; ILP-2; NAIP;

Survivin; Brace; IAPL-3; fortilin; leupeptine; PD- 150606 (3-(4-lodophenyl)-2-mercapto-(Z)-2

propenoic acid); MDL-28170 (Z-Val-Phe-CHO); calpeptin; acetyl-calpastatin; MG 132 (N

[(phenylmethoxy)carbonyl]-L-leucyl-N-[(IS)-I-formyl-3 -methylbutyl]-L-leucinamide); MYODUR; BN 82270 (Ipsen); BN 2204 (Ipsen); AHLi-11 (Quark Pharmaceuticals), an mdm2 protein, pifithrin-a (1-(4

Methylphenyl)-2-(4,5,6,7-tetrahydro-2-imino-3(2H)-benzothiazolyl)ethanone); trans-stilbene; cis

stilbene; resveratrol; piceatannol; rhapontin; deoxyrhapontin; butein; chalcon; isoliquirtigen; butein;

4,2',4'-trihydroxychalcone; 3,4,2',4',6'-pentahydroxychalcone; flavone; morin; fisetin; luteolin;

quercetin; kaempferol; apigenin; gossypetin; myricetin; 6-hydroxyapigenin; 5-hydroxyflavone; 5,7,3

,4, 5'-pentahydroxyflavone; 3,7,3',4',5'-pentahydroxyflavone; 3,6,3',4'-tetrahydroxyflavone; 7,3 ',4

,5 ' -tetrahydroxyflavone; 3 ,6,2 ' ,4 ' -tetrahydroxyflavone; 7,4' -dihydroxyflavone; 7,8,3 ',4'

tetrahydroxyflavone; 3, 6,2',3 '-tetrahydroxyflavone; 4'-hydroxyflavone; 5-hydroxyflavone; 5,4'

dihydroxyflavone; 5,7-dihydroxyflavone; daidzein; genistein; naringenin; flavanone; 3,5,7,3 ',4'

pentahydroxyflavanone; pelargonidin chloride; cyanidin chloride; delphinidin chloride; (-)

epicatechin (Hydroxy Sites: 3,5,7,3',4A; (-)-catechin (Hydroxy Sites: 3,5,7,3 ,4); (-)-gallocatechin

(Hydroxy Sites: 3,5,7,3 ',4',5) (+)-catechin (Hydroxy Sites: 3,5,7,3',4A; (+)-epicatechin (Hydroxy Sites:

3,5,7,3 ',40; Hinokitiol (b-Thujaplicin; 2-hydroxy-4-isopropyl-2,4,6-cycloheptatrien-1-one); L-(+) Ergothioneine ((S)-a-Carboxy-2,3-dihydro-N,N,N-trimethyl-2-thioxo-IH-iniidazole4-ethanaminium

inner salt); Caffeic Acid Phenyl Ester; MCI-186 (3-Methyl-1-phenyl-2-pyrazolin-5-one); HBED (N,N'-Di

(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid*H20); Ambroxol (trans-4-(2-Amino-3,5

dibromobenzylamino)cyclohexane-HCI; and U-83836E ((-)-2-((4-(2,6-di-1-Pyrrolidinyl-4-pyrimidinyl)- 1

-piperzainyl)methyl)-3,4-dihydro-2,5,7,8-tetramethyl-2H- 1 -benzopyran-6-ol»2HCI); -1 -5-methyl

nicotinamide-2'-deoxyribose; /3-D- 1'-5-methyl-nico-tinamide-2'-deoxyribofuranoside; /3-1'-4,5

dimethyl-nicotinamide-2'-de-oxyribose; /3-D-1 '-4,5-dimethyl-nicotmamide-2'-deoxyribofuranoside;

1-Naphthyl PPI (1-(1,1-Dimethylethyl)-3-(I-naphthalenyl)-H-pyrazolo[3, 4-d]pyrimidin-4-amine);

LavendustinA(5-[[(2,5-Dihydroxyphenyl)methyl][(2-hydroxyphenyl)methy 1] amino] -2

hydroxybenzoic acid); MNS (3,4-Methylenedioxy-b-nitrostyrene); PPI (-(l,l-Dimethylethyl)--(4

methylphenyl)-IH- pyrazolo[3, 4-d]pyrimidin-4-amine); PP2 (3-(4-chlorophenyl) I-(1,1-dimethylethyl)

IH-pyrazolo[3,4-d]pyrimidin-4-amine); KX1-004 (Kinex); KX1-005 (Kinex); KX1-136 (Kinex); KX1-174

(Kinex); KX1-141 (Kinex); KX2-328 (Kinex); KX1-306 (Kinex); KX1-329 (Kinex); KX2-391 (Kinex); KX2 377 (Kinex); ZD4190 (Astra Zeneca; N-(4-bromo-2-fluorophenyl)-6-methoxy-7-(2-(IH-1,2,3-triazol-l

yl)ethoxy)quinazolin-4-amine); AP22408 (Ariad Pharmaceuticals); AP23236 (Ariad Pharmaceuticals);

AP23451 (Ariad Pharmaceuticals); AP23464 (Ariad Pharmaceuticals); AZD0530 (Astra Zeneca);

AZM475271 (M475271; Astra Zeneca); Dasatinib (N-(2-choro-6-methylphneyl)-2-(6-(4-(2

hydroxyethyl)-piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide); GN963 (trans

4-(6,7-dimethoxyquinoxalin-2ylamino)cyclohexanol sulfate); Bosutinib (4-((2,4-dichloro-5

methoxyphenyl)amino)-6-methoxy-7-(3-(4-methyl-l-piperazinyl)propoxy)-3-quinolinecarbonitrile); or

combinations thereof.

[00335] In some embodiments, the drug is an agent that reduces undesired neuron or nerve impulses. In some embodiments, the drug reduces one or more symptoms ofdyskinesia or

synkinesia. In some embodiments, the drug is carbamazepine, oxcarbazepine, phenytein, valproic

acid, sodium valproate, cinnarizine, flunarizine, or nimodipine, or combinations thereof.

[00336] In some embodiments, the drug is an agent that promotes regeneration of neuron or nerve tissue. In some embodiments, the drug is a growth factor. In some embodiments, the drug

is selected from brain-derived neurotrophic factor (BDNF); ciliary neurotrophic factor (CNTF); glial

cell-line derived neurotrophic factor (GDNF); neurotrophin-3; neurotrophin-4; fibroblast growth

factor (FGF) receptor; insulin- like growth factor (IGF); or a combination thereof.

XIII. Methods of Light Induced Nerve Ablation

[00337] The present disclosure provides methods of delivering a photosensitizing agent to a human neuron or nerve comprising: contacting the human neuron or nerve with a human neuron or

component of either, and (b) a photosensitizing agent. In some embodiments, the method further

comprises exposing the human neuron or nerve with a light source that activates the

photosensitizing agent, wherein the activated photosensitizing agent induces ablation or killing of

the human neuron or nerve. Upon exposure to a specific wavelength of light, a photosensitizing

agent reacts with molecular oxygen to produce singlet oxygen, which is cytotoxic. In certain

embodiments, a photosensitizing agent is a porphyrin, chlorin, or dye. Examples of photosensitizing agents include porphyrin, protoporfin IX, purlytin, verteporfin, HPPH, temoporfin, methylene blue,

aminileuvolinic acid, Lutetium texaphyrin, metallophthalocyanine, metallo

naphthocyaninesulfobenzo-porphyrazines, metallo-naphthalocyanines, zinc

tetrasulfophthalocyanine, bacteriochlorins, metallochlorins, chlorine derivative, Tetra(m

hydroxyphenyl)chlorin (mTHPC), pheophorbide, dibromofluorescein (DBF), IR700DX,

naphthalocyanine, and porphyrin derivatives. In some embodiments, the human neuron or nerve

targeting molecule comprises a peptide sequence comprising SGQVPWEEPYYVVKKSS (HNP 401; SEQ

ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID

NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with

Ac-DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker;

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20),

SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28),

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), or 5FAM

the peptide comprises: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ

(HNP 402; SEQ ID NO:2), or DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3). In some embodiments the

peptide comprises: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKGGC

(HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC

linker; SEQ ID NO:7), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), QVPWEEPYYVVKKSS

(HNP401-N-2; SEQ ID NO:20), or QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).

In some embodiments the peptide comprises SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In

some embodiments the peptide comprises WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In some

embodiments the peptide comprises DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3). In some

embodiments the peptide comprises Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4). In some embodiments the peptide comprises Ac-WEYHYVDLNWTSQHPQGGC (HNP402

with GGC linker; SEQ ID NO:5). In some embodiments the peptide comprises Ac

DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6). In some embodiments the peptide

comprises QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20). In some embodiments the peptide

comprises QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21). In some embodiments the peptide comprises Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ

ID NO:7). In some embodiments the peptide comprises PWEEPYYVVKKSS (HNP401-N-4; SEQ ID

NO:22). In some embodiments the peptide comprises Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8). In some embodiments the peptide comprises EEPYYVVKKSS (HNP401-N-6;

SEQ ID NO:23). In some embodiments the peptide comprises Ac-EEPYYVVKKSSGGC (HNP401-N-6

with GGC linker; SEQ ID NO:9). In some embodiments the peptide comprises PYYVVKKSS (HNP401-N

8; SEQ ID NO:24). In some embodiments the peptide comprises Ac-PYYVVKKSSGGC (HNP401-N-8

with GGC linker; SEQ ID NO:10). In some embodiments the peptide comprises SGQVPWEEPYYVVKK

(HNP401-C-2; SEQ ID NO:25). In some embodiments the peptide comprises Ac

SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11). In some embodiments the

peptide comprises SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26). In some embodiments the

peptide comprises Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12). In some

embodiments the peptide comprises SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27). In some

embodiments the peptide comprises Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID

NO:13). In some embodiments the peptide comprises SGQVPWEEP (HNP401-C-8; SEQ ID NO:28). In

some embodiments the peptide comprises 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104). In some embodiments the peptide comprises Ac-SGQVPWEEPGGC (HNP401

C-8 with GGC linker; SEQ ID NO:14). In some embodiments the peptide comprises

comprises a peptide that is not Ac-SHSNTQTLAKAPEHTGC (Ac-NP41 with GC linker; SEQ ID NO:17).

In some embodiments, the targeting molecule comprises a peptide that is not SHSNTQTLAKAPEHTGC

(NP41 with GC linker; SEQ ID NO:18). In some embodiments, the targeting molecule comprises a

peptide that is not NTQTLAKAPEHT (SEQ ID NO:19).

[00338] Human neuron or nerve targeting molecules comprising a photosensitizing agent as disclosed herein can be used in methods of localized nerve killing in a subject. In some

embodiments, human neuron or nerve targeting molecules comprising a photosensitizing agent are

used for treating chronic pain (e.g., back, neck, or joint pain) in subject. In some embodiments,

human neuron or nerve targeting molecules comprising a photosensitizing agent are used for

treating prostate cancer in a subject. Autonomic innervation may contribute to prostate cancer

growth and metastasis by light induced ablation of local autonomic nerves. Thus local autonomic

nerves may be a viable target for prostate cancer therapy. In some embodiments, human neuron or

nerve targeting molecules comprising a photosensitizing agent are used for treating renovascular

hypertension in a subject by light induced ablation of sympathetic nerves in the renal vessels. In some embodiments, human neuron or nerve targeting molecules comprising a photosensitizing agent are used for treating excessive sweating. In some embodiments, human neuron or nerve targeting molecules comprising a photosensitizing agent are used for treating cardiac arrhythmias. In some embodiments, human neuron or nerve targeting molecules comprising a photosensitizing agent are used for treating pathological muscle spasms (e.g., Meige syndrome, hemifacial spasm, torticollis).

XIV. Pharmaceutical Compositions

[00339] Disclosed herein, in certain embodiments, are pharmaceutical compositions comprising a human neuron or nerve targeting molecule disclosed herein. Pharmaceutical

compositions herein are formulated using one or more physiologically acceptable carriers including

excipients and auxiliaries which facilitate processing of the active agents into preparations which are

used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. A summary of pharmaceutical compositions is found, for example, in Remington: The Science and

Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E.,

Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.

A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N. Y., 1980; and

Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams

& Wilkins, 1999). In some embodiments, the human neuron or nerve targeting molecule comprises a

peptide sequence selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID

DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC

SEQ ID NO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9), Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC

with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20),

SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28),

with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG

NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), and 5FAM

NO:1), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac

comprises the peptide SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1). In some embodiments, the targeting molecule comprises the peptide WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2). In some

NO:3). In some embodiments, the targeting molecule comprises the peptide Ac

NTQTLAKAPEHT (SEQ ID NO:19).

[00340] In certain embodiments, a pharmaceutical composition disclosed herein further comprises a pharmaceutically acceptable diluent(s), excipient(s), or carrier(s). In some embodiments, the pharmaceutical compositions include other medicinal or pharmaceutical agents, carriers,

adjuvants, such as preserving, stabilizing, wetting or emulsifying agents, solution promoters, salts for

regulating the osmotic pressure, and/or buffers. In addition, the pharmaceutical compositions also

contain other therapeutically valuable substances.

[00341] In certain embodiments, the human neuron or nerve targeting molecules disclosed herein are delivered to a subject via a drug delivery vehicle or carrier. In some embodiments, a

delivery vehicle is made from natural or synthetic materials or both. In some embodiments, a

delivery vehicle is a nanoparticle, microparticle, polymeric micelle, nanocapsule, dendrimer, large

PEG, nanogel, liposome, fullerene, nanostructured lipid carrier, nanoshell, quantum dot, protein

based nanocarriers (e.g., albumin, elastin, gliadin, legumin, zein, soy protein, milk protein, whey

based nanocarriers), organic nanocarrier (e.g., gelatin, dextran, guar gum, chitosan, collagen), polysaccharide based carrier (e.g., dextran, chitosan, pectin), lipid emulsion, or a combination thereof.

[00342] In certain embodiments, a pharmaceutical composition disclosed herein is administered to a subject by any suitable administration route, including but not limited to,

parenteral (intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular, intrathecal,

intravitreal, infusion, or local) administration.

[00343] Formulations suitable for intramuscular, subcutaneous, or intravenous injection include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions,

suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or

dispersions. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents, or vehicles

including water, ethanol, polyols (propyleneglycol, polyethylene-glycol, glycerol, cremophor and the

like), suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as

ethyl oleate. Proper fluidity is maintained, for example, by the use of a coating such as lecithin, by

the maintenance of the required particle size in the case of dispersions, and by the use of

surfactants. Formulations suitable for subcutaneous injection also contain optional additives such as

preserving, wetting, emulsifying, and dispensing agents.

[00344] For intravenous injections, an active agent is optionally formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution,

or physiological saline buffer.

[00345] Parenteral injections optionally involve bolus injection or continuous infusion. Formulations for injection are optionally presented in unit dosage form, e.g., in ampoules or in multi

dose containers, with an added preservative. In some embodiments, the pharmaceutical composition described herein are in a form suitable for parenteral injection as sterile suspensions,

solutions or emulsions in oily or aqueous vehicles, and contain formulatory agents such as

suspending, stabilizing and/or dispersing agents. Pharmaceutical formulations for parenteral

administration include aqueous solutions of an active agent in water soluble form. Additionally,

suspensions are optionally prepared as appropriate oily injection suspensions.

[00346] In some embodiments, the pharmaceutical composition described herein is in unit dosage forms suitable for single administration of precise dosages. In unit dosage form, the

formulation is divided into unit doses containing appropriate quantities of an active agent disclosed herein. In some embodiments, the unit dosage is in the form of a package containing discrete quantities of the formulation. Non-limiting examples are packaged tablets or capsules, and powders in vials or ampoules. In some embodiments, aqueous suspension compositions are packaged in single-dose non-reclosable containers. Alternatively, multiple-dose reclosable containers are used, in which case it is typical to include a preservative in the composition. By way of example only, formulations for parenteral injection are presented in unit dosage form, which include, but are not limited to ampoules, or in multi dose containers, with an added preservative.

[00347] In some embodiments, the human neuron or nerve targeting molecule is administered via systemic intravenous injection into human patients.

EXAMPLES

EXAMPLE 1: PEPTIDES FOR TARGETING HUMAN NERVES AND THEIR USE IN IMAGE GUIDED

SURGERY, DIGNOSTICS AND THERAPEUTIC DELIVERY SUMMARY

[00348] Phage display screens to identify peptides that bind human nerves and could therefore be useful for systemic in-vivo labeling of nerves during fluorescence assisted surgery was

used. Specifically, m13 phage libraries expressing 16 random amino acid sequences on the N

terminus of gill (Creative Biolabs) were processed through selections for binding to freshly resected

or frozen human nerves. In parallel, a newly designed NP41X12+4 library was screened. Each library

was processed through up to 6 binding and wash cycles. Selected phage were additionally selected

for counter-selected for low affinity muscles and fat tissue by preabsobing library, And any high

affinity binder, with tissue prior to positive selection for nerve binding. Sequencing of individual

phage yielded these unique sequences that were highly enriched and therefore higher affinity

relative to the pool of clones: SGQVPWEEPYYVVKKSS (HNP401; SEQ ID NO:1),

WEYHYVDLNWTSQHPQ (HNP402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP403; SEQ ID NO:3) from the

X16 library and DTHAHAKPRVPAFKSV (HNP404; SEQ ID NO:16) from NP41-X12+4 library. Amino acid

sequences derived from sequences of selected phage were chemically synthesized as peptides by

solid-phase synthesis and labeled with fluorescein (FAM) or Cy5 at the C-terminus via a GGC linker

for in-vitro binding to human nerves and in-vivo labeling of rodent nerves. Strong labeling of freshly

sections of human nerves and in-vivo labeled mouse sciatic nerves was shown. Useful labeling occurs between 2-6 hours after intravenous administration and could be visualized using a customized fluorescence dissecting microscope, a Maestro imager from CRI, or a Zeiss Lumar.

[00349] Preservation of peripheral nerves is one of the most important goals of any surgical procedure, because accidental transection of peripheral nerves during surgical procedures lead to

significant morbidity for patients. Also, nerves grow back more slowly and incompletely after

transection than almost any other tissue. Typically, peripheral nerves are identified by their

relatively constant relationship to nearby structures as well as by their typical appearance of being

elongated whitish, glistening structures. However, in many instances, identification of peripheral nerves using these criteria can be difficult: for example in cases of tumor involvement, in instances

of inflammation/infection, in a previously operated surgical field, or when the nerve is encased in

bone.

[00350] Current methods for nerve labeling primarily depend on retrograde or anterograde tracing of individually identified axonal tracts via the use of fluorescent dyes. The fluorescent dyes

are either applied to the innervation target and travel in a retrograde fashion to label the innervating

nerve fibers, or are applied directly to the identified nerves and label the nerve fibers both

anterogradely and retrogradely. This technique has the drawback of being able to label only one

nerve fiber tract at a time, depending on where the dye has been injected. A second drawback is the

limited accumulation of fluorescent dyes along the axonal tracts, because retrograde axonal tracers

typically accumulate in the neural cell body and axonal labeling with these fluorescent dyes is

limited. A third disadvantage of this technique is that retrograde transport is relatively slow (on the

order of millimeters per day) and therefore takes a long time to label human nerves, which are often

longer than a meter (as in the case of the sciatic nerve and its arborizations). Furthermore, the

application of fluorescent dyes to innervation targets such as direct intramuscular injections to label

motor nerves is typically messy with a variable amount of the tracer dye remaining at the injection

site. As dissection of nerves depends on accurate visualization of adjacent structures prior to

encountering them, a surgical site that is contaminated with fluorescent dyes would not be

desirable. Finally, the direct injection of the fluorescent dye itself may be damaging to the target

organs or nerve of interest, either by mechanical damage or by the very high local concentration of

dye and vehicle at the injection site.

[00351] The method of systemic injection of fluorescently labeled peptides to label nerves described in this document addresses all of the disadvantages of fluorescently tracers described above. First, as the peptides are delivered systemically, all peripheral nerves in the body have the potential of being labeled. This is contrast to the labeling of only one nerve at a time as with current methods. Second, as the peptides described here were selected for their ability to bind nerves, the nerve fibers are clearly visualized compared to adjacent non-neural structures. This is in contrast to the preferential accumulation to neural cell bodies rather than axonal processes with most current fluorescent dyes. Third, the binding of the peptides described here to nerves occurs very quickly and visualization of peripheral nerves using this technique can be accomplished within hours. This is in contrast to the relatively slow rate of labeling with anterograde or retrograde tracers. Finally, since the peptides are applied systemically via intravenous injection, damage to nerves at the injection site is not an issue.

[00352] Nerve-homing peptide sequences that were derived using mouse peripheral nerves for laboratory research have been previously described (U.S. Patent No. 8,685,372, April 1s' 2014).

However, because the intended eventual clinical application of nerve labeling is in human patients,

identification of unique peptide sequences that bind human nerves was sought. The peptide

sequences described in this application was identified by their ability to bind human nerves. These

peptide sequences were identified by their ability to bind human nerves, following systemic

intravenous injection into human patients, and as such these peptides will be much more likely to

bind human nerves compared to sequences that were selected against rodent nerves.

[00353] Current methods for labeling nerves involve the application of fluorescent tracer dyes (Fast Blue, Rhodamine-isothiocyanate, Fluoro-Ruby, Fluoro-Emerald), carbocyanine dyes (Dil,

DiAsp, DiO, DiA), Fluoro-Gold, fluorescently labeled latex beads, fluorescently labeled plant lectins

and bacterial toxins (wheat germ agglutinin, peanut agglutinin, concanavalin A, Phaseolus vulgaris

leucoagglutininin (PHA-L), soybean agglutinin, Ulex europaeus agglutinin, Ricinus communis

agglutinin (I and II), tetanus toxin fragment C, cholera toxin B and fluorescently labeled dextran

conjugates.

METHODS:

Experimental details:

[00354] m13 phage libraries expressing random 16 amino acid sequences on the N-terminus of gill (Creative Biolabs) or an internally derived library expressing derivative of NP41 were used to

identify peptides that bond human nerve tissue.

Selection of peptides

[00355] Human peripheral nerves were obtained from patients undergoing nerve resection procedures and homogenized. Phage library mixture was incubated with nerve homogenate or

nerve homogenates that had been bound to high protein binding 6 well plates. Following incubation,

the mixture were either centrifuged and the pellet washed with PBS, or plate was washed with PBS.

The pellet was rehomogenized and plated for titer and re-amplification or released from plate with

low pH buffer. Phage that were bound at each round were sequenced and repeats noted. No

repeats were identified until round 4 of selection.

SGQVPWEEPYYVVKKSS (HNP401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP402; SEQ ID NO:2),

DLPDIIWDFNWETA (HNP403; SEQ ID NO:3) from the X16 library and DTHAHAKPRVPAFKSV (SEQ ID

NO:16) were identified after round 5, Table 1

Table 1: Peptides Identified

Name Repeating sequences Number of SEQ ID NO: repeats

HNP 401 SGQVPWEEPYYVVKKSS >10 1

HNP 402 WEYHYVDLNWTSQHPQ >10 2

HNP 403 DLPDIIWDFNWETA 7 3

HNP404 DTHAHAKPRVPAFKSV 3 16

In vivo testing of peptides

[00356] Either 150 or 450 nmoles nmoles of fluorescein labeled synthetic peptides were injected intravenously into mice. After a 2 hour waiting period for washout of nonspecific binding,

mice were anesthetized and skin incisions were made over the dorsal surface of the hind legs to

expose the sciatic nerve. Brightfield and fluorescent images were obtained with a dissecting

microscope using Metamorph software (Figure 1). Quantitation of fluorescence of nerves and

adjacent non-nerve tissue was performed with Image J (Figure 2). Peptides were also topically

applied to human nerve sections. Nerves were freshly frozen in OTC prior to sectioning. Peptides

were topically applied at concentration at 300uM with images being shown for HNP401, HNP402,

HNP404 and previous reported nerve binding peptides NP41 (Figure 3)). Also shown are images for

variants HNP301 (SEFPRSWDMETN) and NP124. NP713 was also tested and has not not reported in a publication. NP713, is a derivative of NP41 with sequence NTHPHTTSRVPSQIAR that was enriched after 7 rounds of selection against mouse tissue, and was also found after 4 rounds of selection against human tissue. Binding of NP713 phage compared to wildtype phage showed a 4.8-fold higher nerve:muscle ratio. FAM-NP713 showed similar nerve:muscle contrast to NP41 (Data not shown here). All D-amino acid controls for NP-41 and NP713 and non peptide conjugated carboxyfluorescein, are also shown. HNP401 shows the highest nerve specific contrast with the majority of the labelling occurring in the perineurium. To further demonstrate HNP401 selective binding, HNP401, NP41 and HNP404 were tested at a lower concentration, 100.IM (Figure 4).

HNP401 was then tested for labeling of rat sciatic and rat prostate cavernosal nerve in-vivo. Figure 5

shows in-vivo labeling of rat sciatic nerve. Figures 6 and 7 show in-vivo labeling of rat prostate

cavernosal nerve with comparison to white light visualization.

[00357] Nerve-homing peptides sequences that were identified by their ability to bind mouse nerves for laboratory research were previously described. Because the peptide sequences

described in this document were identified by their ability to bind human nerves, following systemic

intravenous injection into human patients, these peptides will be much more likely to bind human

nerves compared to sequences that were selected against rodent nerves.

[00358] Fluorescently labeled human nerve-binding peptides are applied systemically via intravenous injection. Following a short waiting period for washout of nonspecific binding,

peripheral nerves can be visualized within a surgical field with appropriate excitation and emission

filters.

[00359] Human nerve-binding peptides might also be conjugated to factors that may have neurotrophic or protective properties to nerves. Following systemic application via intravenous

injection, peptide-trophic/neuroprotective factor conjugates might facilitate repair/regeneration of

damaged nerves both in the periphery and in the spinal cord.

[00360] Human nerve binding peptides conjugated to neuroprotective/neurotrophic factors may also be conjugated to injury homing peptides to further improve localized delivery of these

factors to injured nerves, potentially faciliting resistance to injury/repair/regeneration.

Applications & Uses:

[00361] Fluorescently labeled nerve-binding peptides can be used to assist surgeons in the visualization of nerves during surgical procedures prior to physically encountering and thus

potentially damaging them. This is particularly important during surgery on the prostate gland,

because the cavernosal nerves controlling male erections run very near the prostate but are

practically invisible ordinarily.

[00362] Nerve binding peptide-neurotrophic/neuroprotective factor conjugates can be used to facilitate repair/regeneration of damaged nerves.

[00363] Nerve binding peptides could be conjugated to photosensitizing dyes for potential use with light induced nerve killing as a treatment for localized pain

REFERENCES:

1. Whitney M, Crisp J, Nguyen L, Friedman B, Gross L, Steinbach P, Tsien R, Nguyen Q.

Fluorescentpeptides highlight peripheral nerves during surgery in mice. Nature Biotechnology. 2011;29:352-356

2. Wu AP, Whitney MA, Crisp JL, Friedman B, Tsien RY, Nguyen QT. Improved facial nerve

identification with novel fluorescently labeled probe. The Laryngoscope. 2011;121:805-810

3. Kobbert C., Apps, R., Bechmann, I., Laciego, J.L., Mey, J., Thanos, S. Currents concepts in

neuroanatomical tracing. Progress in Neurobiology 62 (2000) 327-351.

4. Richmond, F.J.R., Gladdy R., Creasy, J.L., Ktamura S., Smits, E., Thomson D.B. Efficacy of seven

retrograde tracers, compared in multiple-labelling studies of feline motoneurones. Journal of

Neuroscience Methods 53 (1994) 35-46.2

5. Marangos, N., Illing R., Kruger J., Laszig R. In vivo visualization of the cochlear nerve and nuclei

with fluorescent axonal tracers. Hearing Research 162 (2001) 48-52.

6. O'Malley, M, Wittkopf, J., Cutler J., Labadie, R, Hackett, T, Haynes, D. Fluorescent retrograde

axonal tracing of the facial nerve. The Laryngoscope 116 (2006) 1792-1797.

EXAMPLE 2: OPTIMIZED PEPTIDES FOR TARGETING HUMAN NERVES AND THEIR USE IN IMAGE GUIDED SURGERY, DIGNOSTICS AND THERAPEUTIC DELIVERY

SUMMARY

[00364] Phage display screens were used to identify peptides that bind human nerves and could therefore be useful for systemic in-vivo labeling of nerves during fluorescence assisted surgery. Specifically, m13 phage libraries expressing 16 random amino acid sequences on the N-terminus of gill (Creative Biolabs) were processed through selections for binding to freshly resected or frozen human nerves. Library was processed through up to 6 binding and wash cycles. Selected phage were additionally selected for counter-selected for low affinity muscles and fat tissue by preabsobing library, And any high affinity binder, with tissue prior to positive selection for nerve binding. Sequencing of individual phage yielded these unique sequences that were highly enriched and therefore higher affinity relative to the pool of clones: SGQVPWEEPYYVVKKSS (HNP401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP403; SEQ ID NO:3). Amino acid sequences derived from sequences of selected phage were chemically synthesized as peptides by solid-phase synthesis and labeled with fluorescein or Cy5 for in-vitro binding to human nerves and in-vivo labeling of rodent nerves. Strong labeling of freshly sections of human peripheral nerves (motor, sensory, autonomic) and in-vivo labeled mouse and rat sciatic nerves was shown. Useful labeling occurs between 2-6 hours after intravenous administration and could be visualized using a customized fluorescence dissecting microscope, a Maestro imager from CRI, or a Zeiss Lumar dissecting microscope.

RESULTS

Nerve identificationand preservation is essential in surgery of the head and neck.

[00365] Identification of peripheral nerves is critical for their preservation during surgery, because accidental transection or injury can lead to significant patient morbidity including chronic pain, numbness or permanent paralysis'. Nerve identification is especially important during surgery of the head and neck. For example, facial nerve dysfunction has been reported to be as high as 40% during the acute postoperative period and 30% at1 month following parotidectomy . Similarly, facial nerve dysfunction has been reported to be as high as 30% atyear following vestibular schwannoma surgery 4. Temporary and permanent vocal fold immobility are major surgical complications of thyroid surgery, anterior cervical approaches to the spine, esophagectomy and carotid endarterectomy '. Although the course of the facial nerve typically follow defined anatomical landmarks, extensive patient to patient variability has been documented for every branch of the extratemporal facial nerve including variability in total number of divisions, origin of individual divisions and connections between divisions 6. Even within the same patient, the left and right facial nerve may display differences in course and divisions ". Similar variability has been documented for the recurrent laryngeal nerve ". In instances of tumor invasion, inflammation, trauma or repeat surgery, nerve identification can be even more challenging. Finally, identification of degenerated nerves, which are critically important during reconstructive surgery, is even more difficult than their functioning counterpart as they become smaller and thinner over time.

Consequently, any means of improving the visual determination between nerve and non-nerve

tissue would represent a significant advance in surgical technique.

Nerve identificationand preservation is essential during other surgeries includingprostate cancer

surgery.

[00366] Prostate cancer is the most common solid organ malignancy in U.S. men. For men with localized prostate cancer surgery results in excellent cancer control. All too often this cancer

treatment comes at the expense of erectile function, urinary control, and overall quality of life. Preservation of the autonomic neurovascular bundles during radical prostatectomy is an important

aspect of the operation. For nearly 20 years the importance of preserving the autonomic nerves that

run along the posterolateral aspect of the prostate in order to preserve erectile function has been

recognized. The autonomic nerve fibers themselves are rarely visualized, however. Instead

surgeons preserve the blood vessel complexes, or neurovascular bundles, that have been shown to

have the highest density of autonomic nerves. The exact position and distribution of these

autonomic nerves are variable 13-18as even in the most experienced hands, erectile dysfunction and

urinary incontinence are common'. Improved sexual function outcomes are associated with

increasing surgeon experience and avoidance of crush or traction injuries on these nerves.

Significant anatomic variation and differences in surgeon experience and volume create an

opportunity to improve surgical quality while minimizing adverse outcomes. In instances of tumor

invasion, inflammation, trauma or repeat surgery, nerve identification and preservation would

represent an additional challenge. Finally, the growing use of robotically-assisted surgery, with its inherent lack of haptic feedback , further increases the surgeon's dependence on visual

information. Consequently, any means of improving the visual determination between nerve and

non-nerve tissue would represent a significant advance.

Small nerves are hard to identifyduring surgery.

[00367] Thin or buried nerves are particularly difficult to distinguish and are therefore the most likely to be damaged during surgical procedures. Identification of motor nerves prior to direct exposure is currently dependent on electromyographic (EMG) monitoring20-22, in which a stimulating electrode is inserted and distal muscle twitches are monitored. EMG is not an imaging technique, so even if a nerve has been identified in one location there is no visual guidance as to how far from the stimulation site and in which direction the nerve lies. Furthermore, EMG only identifies motor pathways, not sensory fibers such as the first two divisions of the trigeminal nerve or the cochleovestibular nerve, nor sympathetic tracts such as the neurovascular bundle surrounding the prostate gland 23-25 , where nerve injury following radical prostatectomy leads to significant urinary incontinence and erectile dysfunction ". Electrode insertion may itself damage a nerve. Finally, EMG fails if axonal or neuromuscular transmission is temporarily blocked distal to the recording site by nerve compression, trauma, tumor invasion, local anesthetics, or neuromuscular blockers. There are some potential technologies for in vivo nerve visualization without exogenous probe molecules, such as optical coherence tomography or laser confocal microscopy28. However, nerves have very little intrinsic contrast to distinguish them from other tissues, and these techniques do not readily produce real-time live images over the field of view necessary for guiding surgery. Degenerated nerves, important to identify during reconstructive surgery after cancer resection, traumatic or therapeutic amputations, would also have no myelin and therefore would not benefit from these agents.

Competing strategies to improve nerve visualizationduring surgery.

[00368] For these reasons, there is much interest in development of labeling reagents to improve nerve visualization during surgery. There has been focus on nerve labeling which depend

on retrograde or anterograde tracing of individually axonal tracts via the use of fluorescent dyes -32

or the B subunit of Cholera toxin (CTb488) 33 . The fluorescent dyes are either applied to the

innervation target and travel in a retrograde fashion to label the innervating nerve fibers, or are

applied directly to the identified nerves and label the nerve fibers both anterogradely and

retrogradely. Local injections have the drawback of being able to label only one nerve fiber tract at a time. Anterograde and retrograde transport is relatively slow and can take days to travel a few

millimeters, while leaving most of the tracer at the injection site. As dissection of nerves depends on

accurate visualization of adjacent structures, a surgical site that is heavily contaminated with excess

fluorescent dyes would not be desirable. Finally, the direct injection of the fluorescent dye may be

damaging to the target organs or nerve of interest, either by mechanical damage or by the very high

local concentration of dye and vehicle at the injection site.

[00369] More recently, there has interest in using vascular dyes such as indocyanine green (ICG) to label the vascular supply of nerves (i.e. vaso nervorum) 3 4, 3s. One limitation of this technology is that small nerves (such as cavernosal nerves important for prostate surgery) will have proportionally less vaso nervorum, limiting contrast and intensity compared to adjacent tissue.

[00370] There has also been focus on agents targeting myelin including distyrylbenzene (DSB) derivatives3, coumarin derivatives and anti-ganglioside antibodies 3 . DSB and coumarin derivatives are small molecules with intrinsic fluorescence while anti-ganglioside antibodies are conjugated to fluorescent dyes for imaging 3s- 43. While these molecules are potentially promising for peripheral nerve imaging, non-myelinated nerves such as cavernosal nerves (which are autonomic and minimally myelinated) would likely have little binding, thereby limiting their utility in these important surgeries. Degenerated nerves would also have limited myelin present and thus would not be labeled with these agents.

[00371] The method of systemic injection of fluorescently labeled peptides to label nerves described in this document addresses all of the disadvantages of other nerve targeting techniques described above. First, as the peptides are delivered systemically, all peripheral nerves in the body have the potential of being labeled. This is contrast to the labeling of only one nerve at a time as with current methods. Second, as the peptides described here were selected for their ability to bind nerves, the nerve fibers are clearly visualized compared to adjacent non-neural structures. This is in contrast to the preferential accumulation to neural cell bodies rather than axonal processes with most current fluorescent dyes. Third, the binding of the peptides described here to nerves occurs very quickly and visualization of peripheral nerves using this technique can be accomplished within hours. This is in contrast to the relatively slow rate of labeling with anterograde or retrograde tracers. Finally, since the peptides are applied systemically via intravenous injection, damage to nerves at the injection site is not an issue.

[00372] Nerve-homing peptide sequences that were derived using mouse peripheral nerves for laboratory research have been previously described (U.S. patent 8,685,372, April 1st 2014 Peptides and aptamers for targeting of neuron or nerves US20120148499 and W2010121023A2). However, because the intended eventual clinical application of nerve labeling is in human patients, identification of unique peptide sequences that bind human nerves was sought. The peptide sequences described in this application was identified by their ability to bind human nerves. These peptide sequences were identified by their ability to bind human nerves, following systemic intravenous injection into human patients, and as such will be much more likely to bind human nerves compared to sequences than peptides that were selected against rodent nerves.

[00373] Current methods for labeling nerves involve the application of fluorescent tracer dyes (Fast Blue, Rhodamine-isothiocyanate, Fluoro-Ruby, Fluoro-Emerald), carbocyanine dyes (Dil,

agglutinin (I and II), tetanus toxin fragment C, cholera toxin B and fluorescently labeled dextran conjugates.

[00374] The fluorescent dyes are either applied to the innervation target and travel in a retrograde fashion to label the innervating nerve fibers, or are applied directly to the identified

nerves and label the nerve fibers both anterogradely and retrogradely. As mentioned above, local

injections have the drawback of being able to label only one nerve fiber tract at a time. Anterograde

and retrograde transport is relatively slow and can take days to travel a few millimeters, while

leaving most of the tracer at the injection site. As dissection of nerves depends on accurate

visualization of adjacent structures, a surgical site that is heavily contaminated with excess

local concentration of dye and vehicle at the injection site.

[00375] More recently, there has interest in using vascular dyes such as indocyanine green (ICG) to label the vascular supply of nerves (i.e. vaso nervorum) 3 4, 3s. One limitation of this

technology is that small nerves (such as cavernosal nerves important for prostate surgery) will have

proportionally less vaso nervorum, limiting contrast and intensity compared to adjacent tissue.

[00376] There has also been focus on agents targeting myelin including distyrylbenzene (DSB) derivatives3, coumarin derivatives and anti-ganglioside antibodies 3 . DSB and coumarin

derivatives are small molecules with intrinsic fluorescence while anti-ganglioside antibodies are

conjugated to fluorescent dyes for imaging 36- 43 . While these molecules are potentially promising for

peripheral nerve imaging, non-myelinated nerves such as cavernosal nerves (which are autonomic

and minimally myelinated) would likely have little binding, thereby limiting their utility in these

important surgeries. Degenerated nerves would also have limited myelin present and thus would

not be labeled with these agents.

[00377] Nerve-homing peptides sequences that were identified by their ability to bind mouse nerves for laboratory research were previously described. Because the peptide sequences described in this document were identified by their ability to bind human nerves, following systemic intravenous injection into human patients, these peptides will be much more likely to bind human nerves compared to sequences that were selected against rodent nerves.

[00378] Fluorescently labeled human nerve-binding peptides are applied systemically via intravenous injection. Following a short waiting period for washout of nonspecific binding, peripheral nerves can be visualized within a surgical field with appropriate excitation and emission filters.

[00379] Human nerve-binding peptides might also be conjugated to factors that may have neurotrophic or protective properties to nerves. Following systemic application via intravenous injection, peptide-trophic/neuroprotective factor conjugates might facilitate repair/regeneration of damaged nerves both in the periphery and in the spinal cord.

[00380] Human nerve binding peptides conjugated to neuroprotective/neurotrophic factors may also be conjugated to injury homing peptides to further improve localized delivery of these factors to injured nerves, potentially faciliting resistance to injury/repair/regeneration.

METHODS

Experimental details:

[00381] m13 phage libraries expressing random 16 amino acid sequences on the N-terminus of gill (Creative Biolabs) or an internally derived library expressing derivative of NP41 to identify peptides that bond human nerve tissue were used. NTQTLAKAPEHT (NP-41; SEQ ID NO:15; see U.S. Patent No. 8,685,372 or International Patent Publication No. W02010121023A2; both of which are incorporated by reference herein in there entireties).

Selection of peptides:

[00382] Human peripheral nerves were obtained from patients undergoing nerve resection procedures and homogenized. The phage library mixture was incubated with nerve homogenate or nerve homogenates that had been bound to high protein binding 6 well plates. Following incubation, the mixtures were either centrifuged and the pellet washed with PBS, or plate was washed with PBS.

repeats were identified until round 4 of selection.

[00383] The following peptides were identified: SGQVPWEEPYYVVKKSS (HNP401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP403; SEQ ID NO:3)

from the X16 library and DTHAHAKPRVPAFKSV (SEQ ID NO:16) were identified after round 5. See

peptides Table 2.

Table 2: Peptides Identified

Name Repeating sequences Number of SEQ ID NO: repeats

HNP 401 SGQVPWEEPYYVVKKSS >10 1

HNP 402 WEYHYVDLNWTSQHPQ >10 2

HNP 403 DLPDIIWDFNWETA 7 3

Demontration of peptide binding to human nerves:

[00384] To determine the affinity of phage selected peptides for binding to human nerve they were chemically synthesized by solid-phase synthesis and labeled with fluorescein at the C

terminus via a GGC linker. Peptides were topically applied to sectioned human sural nerve and

human temporalis muscle to determine nerve to muscle contrast. HNP401 showed the highest

binding and highlighting of human never (Figure 8). Data for other peptides screened on human

nerves and controls including free carboxy fluorescein and NP41 screened on human nerve are

additionally shown. To confirm binding and contrast of HNP401 for additional nerve types binding

was compared in nerve and muscule with both HNP401 and NP41 on facial brachial plexus nerve

(Figure 9). To quantify differential binding to human nerve versus muscle, fluorescence signal

intensity was measured for ROls from the perineurium of select nerves and human temporalis

muscle that had been identically treated with topical application of fluorescein labelled nerve

binding peptides. FAM-HNP401 showed selective binding to human sural nerve with 10.9X

fluorescent signal intensity (1374.44 ±425.96) compared to FAM-NP41 (126.17 ± 61.03) (Fig 9G),

p=0.009, Student's t-test, unpaired). Nerve to muscle contrast was comparable at 3.03 i 0.57 for

FAM-HNP401 and 2.28 i 0.96 for FAM-NP41 (Fig 9H), p=0.236, Student's t-test, unpaired). Dose

dependent testing shows HNP401 has significant nerve binding down to 10uM (Figure 10A-E) with increased nerve human binding of HNP401 compared to NP41 detected at concentrations as high

375uM (Figure 10 F-I). FAM-HNP401 was additionally tested topically on ex-vivo tissue for labeling of

mouse facial nerve with surrounding muscle (Figure 10, J-M). Confocal imaging also showed that HNP401-FAM binds epineurium, perineurium and endonerium but not axons (Figure ION).

[00385] In-vivo fluorescence imaging of sciatic nerve in mice that were injected with 450nmols of FAM-HNP401(Figure 11A) or FAM-NP41(Figure 11B) showed nerve contrast with

HNP401 having 2.3 fold fluorescent intensity compared to (NP41) (Figure IC) but with similar nerve

to non-nerve contrast 5.79 ±0.81 for FAM-HNP401 and 6.63 ±1.63 for FAM-NP41 (Figure iD). FAM-HNP401 also highlighted rat prostate nerve (Figure 11 E-F) and rat sciatic nerve (Figure 11G) at

a dose of 2ptmole when imaged 3 hours post injection or alternatively using a lower dose of

0.5.tmole HNP401 with imaging 10 mins after probe injection. Blood clearance of FAM-HNP401 after

injection of 100nmols i.v. showed a half-life of 30 minutes which is similar to FAM-NP41 (Figure

11H).

[00386] FAM-HNP401 and FAM-NP41 were then tested topically for binding to autonomic nerves, isolated from excised prostate glands of two human patients (Figure 12 and 13). FAM

HNP401 (Figure 12A) showed a significantly higher fluorescent signal in human autonomic nerves

compared to FAM-NP41(Figure 12B) at the same concentration. Labelled fiber were confirmed as

nerve using anti-neurofilament antibody SM1312 (red) and DAPI (blue) to show nuclear labeling

(Figure 12C). H&E staining was also done to confirm label tissue as nerve (Figure 12D). Prostate

nerve binding of HNP401 using tissue from an additional patient is shown in Figure 13. Similar

staining was obtained for peripheral sensory anti-brachial cutaneous nerve isolated from a human

arm (Figure 12E-H).

Synthesis and nerve binding of deletion variants of HNP401

[00387] To optimize the HNP401 sequence, systematic deletion of 2 amino acids from the C or N terminus (Table 3) followed by binding analysis to human nerve sections was performed.

Deletion of amino acids from C terminal reduces binding efficacy and solubility. Removal of 2 amino

acids from the N-termini improves the nerve binding with an average signal intensity of 1498.73 (+/

517.63) for N-2 and 744.63 (+/- 130.18) for HNP401 [Student's t test, unpaired, 1 tail, p=0.07] (Figure

14 and 15).

Table 3. List of nerve binding peptides

Unique Peptide Peptide sequence SEQ ID NO:

HNP401 (with GGC linker) Ac-SGQVPWEEPYYVVKKSSGGC 4

HNP401 (with GGC linker) Ac-WEYHYVDLNWTSQHPQGGC 5

HNP403 (with GGC linker) Ac-DLPDIIWDFNWETAGGC 6

HNP401-N-2 (with GGC linker) Ac-QVPWEEPYYVVKKSSGGC 7

HNP4O1-N-4 (with GGC linker) Ac-PWEEPYYVVKKSSGGC 8

HNP4O1-N-6 (with GGC linker) Ac-EEPYYVVKKSSGGC 9

HNP4O1-N-8 (with GGC linker) Ac-PYYVVKKSSGGC 10

HNP401-C-2 (with GGC linker) Ac-SGQVPWEEPYYVVKKGGC1

HNP4O1-C-4 (with GGC linker) Ac-SGQVPWEEPYYVVGGC 12

HNP4O1-C-6 (with GGC linker) Ac-SGQVPWEEPYYGGC 13

HNP4O1-C-8 (with GGC linker) Ac-SGQVPWEEPGGC 14

Applications & Uses:

[00388] Fluorescently labeled human nerve-binding peptides can be used to assist surgeons in the visualization of nerves during surgical procedures prior to physically encountering and thus

potentially damaging them. This is particularly important when nerves are small, degenerated,

invaded by cancer, injured by trauma or infection. For example, during surgery on the prostate

gland, the cavernosal nerves controlling male erections run very near the prostate gland but are not

definitively identified using convential light (white light reflectance) available in operating theaters.

[00389] Human nerve binding peptide-neurotrophic/neuroprotective factor conjugates can be used to facilitate repair/regeneration of damaged nerves.

[00390] Human nerve binding peptides could be conjugated to photosensitizing dyes for potential use with light induced nerve killing as a treatment for chronic pain.

[00391] Human nerve binding peptides could be conjugated to photosensitizing dyes for potential use with light induced nerve killing as a treatment for excessive sweating.

[00392] Human nerve binding peptides could be conjugated to photosensitizing dyes for potential use with light induced nerve killing as a treatment for renovascular hypertension.

[00393] Human nerve binding peptides could be conjugated to photosensitizing dyes for potential use with light induced nerve killing as a treatment for cardiac arrhythmias.

[00394] Human nerve binding peptides could be conjugated to photosensitizing dyes for potential use with light induced nerve killing as a treatment for pathologic muscle spasms (Meige

syndrome, hemifacial spasm, torticollis).

REFERENCES:

1. Burke S and Shorten GD. When pain after surgery doesn't go away. Biochem Soc Trans. 2009;37:318-22.

2. Gaillard C, Perie S, Susini B and St Guily JL. Facial nerve dysfunction after parotidectomy: the role of local factors. The Laryngoscope. 2005;115:287-91.

3. Witt RL. Comparing the long-term outcome of immediate postoperative facial nerve dysfunction and vocal fold immobility after parotid and thyroid surgery. J Voice. 2006;20:461-5.

4. Darrouzet V, Martel J, Enee V, Bebear J and Guerin J. Vestibular schwannoma surgery outcomes: our multidisciplinary experience in 400 cases over 17 years. Laryngoscope. 2004;114:681 8.

5. Rosenthal L, Benninger M and Deeb R. Vocal fold immobility: a longitudinal analysis of etiology over 20 years. Laryngoscope. 2007;117:1864-70.

6. Nason RW, Binahmed A, Torchia MG and Thliversis J. Clinical observations of the anatomy and function of the marginal mandibular nerve. IntJ Oral Maxillofac Surg. 2007;36:712-5.

7. Woltmann M, Faveri R and Sgrott EA. Anatomosurgical study of the marginal mandibular branch of the facial nerve for submandibular surgical approach. Braz DentJ. 2006;17:71-4.

8. Lineaweaver W, Rhoton A and Habal M. Microsurgical anatomy of the facial nerve. J Craniofac Surg 1997;8:6-10.

9. Gosain A, Sewall S and Yousif N. The temporal branch of the facial nerve: how reliably can we predict its path? Plast Reconstr Surg 1997;99:1224-33.

10. Tzafetta K and Terzis J. Essays on the facial nerve: Part 1. Microanatomy. Plast Reconstr Surg. 2010;125:879-89.

11. Haller J, Iwanik M and Shen F. Clinically Relevant Anatomy of Recurrent Laryngeal Nerve. Spine (Phila Pa 1976) 2011;Epub ahead of print.

12. Ebraheim N, Lu J, Skie M, Heck B and Yeasting R. Vulnerability of the recurrent laryngeal nerve in the anterior approach to the lower cervical spine. Spine (Phila Pa1976). 1997;15:2664-7.

13. Walsh P. Anatomic radical prostatectomy: evolution of the surgical technique. J Urology. 1998;160:2418-24.

14. Tewari A, Peabody J, Fischer M, Sarle R, Vallancien G, Delmas V, Hassan M, Bansal A, Hemal A, Guillonneau B and Menon M. An operative and anatomic study to help in nerve sparing during laparoscopic and robotic radical prostatectomy. Eur Urology. 2003;43:444-454.

15. Costello A, Brooks M and Cole 0. Anatomical studies of the neurovascular bundle and cavernosal nerves. British Journal of Urology International 2004;94:1071-6.

16. Kalyana CN, Rupesh R, Ashok A and Craig DZ. Nerve-Sparing Surgery Significantly Affects Long-Term Continence After Radical Prostatectomy. Urology. 2007;70:1127-30.

17. Gallina A, Briganti A, Suardi N, Capitanio U, Abdollah F, Zanni G, Salonia A, Rigatti P and Montorsi F. Surgery and erectile dysfunction. Archives Esp Urology. 2010;63:640-8.

18. Zhao L, Meeks J and Nadler R. Robotics in urologic surgery. Minerva Urol Nefrol. 2009;61:331-9.

19. van der Meijden 0 and Schijven M. The value of haptic feedback in conventional and robot assisted minimal invasive surgery and virtual reality training: a current review. Surg Endosc. 2009;23:1180-90.

20. Gantz B. Intraoperative facial nerve monitoring. The American Journal of Otology. 1985;Nov: Suppl:58-61.

21. Miller M and Spiegel J. Identification and monitoring of the recurrent laryngeal nerve during thyroidectomy. Surgical Oncology Clinics of North America. 2008;17:121-44, viii-ix.

22. WE D, JL R and J T. Recurrent laryngeal nerve localization using a microlaryngeal electrode. Otolaryngology Head and Neck Surgery. 1979;87:330-333.

23. J W, AL B, AJ C, JA E, M G, B G, M M, F M, RP M, B R and A V. A critical analysis of the current knowledge of surgical anatomy related to optimization of cancer control and preservation of continence and erection in candidates for radical prostatectomy. European Urology. 2010;57:179-92.

24. J W, M G and H H. Basic principles of anatomy for optimal surgical treatment of prostate cancer. WorldJournal of Urology. 2007;25:31-8.

25. HR K, TY T, L S, J V, MJ H and P D. Impact of nerve sparing technique on patient self-assessed outcomes after radical perineal prostatectomy. Journal of Urology. 2007;178:488-92.

26. Kubler HR, Tseng TY, Sun L, Vieweg J, Harris MJ and Dahm P. Impact of nerve sparing technique on patient self-assessed outcomes after radical perineal prostatectomy. J Urol. 2007;178:488-92; discussion 492.

27. Chitchian S, Weldon T, Fiddy M and Fried N. Combined image-processing algorithms for improved optical coherence tomography of prostate nerves. J Biomed Opt. 2010;15:046014.

28. Zhivov A, Blum M, Guthoff R and Stachs 0. Real-time mapping of the subepithelial nerve plexus by in vivo confocal laser scanning microscopy. BrJ Ophthalmol 2010;94:1133-5.

29. M OM, J W, J C, R L, T H and D H. Fluorescent retrograde axonal tracing of the facial nerve. Laryngoscope. 2006;116:1792-1797.

30. Marangos N, Illing R, KrOger J and Laszig R. In vivo visualization of the cochlear nerve and nuclei with fluorescent axonal tracers. Hearing Research. 2001;162:48-52

31. C K, R A, I B, JL L, J M and S T. Current concepts in neuroanatomical tracing. Progress in Neurobiology. 2000;62:327-351.

32. FJR R, R G, JL C, S K, E S and DB T. Efficacy of seven retrograde tracers, compared in multiple labelling studies of feline motoneurones. Journal of Neuroscience Methods 1994;53:35-46.

33. Boyette LB, Reardon MA, Mirelman AJ, Kirkley TD, Lysiak JJ, Tuttle JB and Steers WD. Fiberoptic imaging of cavernous nerves in vivo. J Urol. 2007;178:2694-700.

34. Wagner OJ, Louie BE, Vallieres E, Aye RW and Farivar AS. Near-infrared fluorescence imaging can help identify the contralateral phrenic nerve during robotic thymectomy. Ann Thorac Surg. 2012;94:622-5.

35. Chen SC, Wang MC, Wang WH, Lee CC, Yang TF, Lin CF, Wang JT, Liao CH, Chang CC, Chen MH, Shih YH and Hsu SP. Fluorescence-assisted visualization of facial nerve during mastoidectomy: A novel technique for preventing iatrogenic facial paralysis. Auris Nasus Larynx. 2015;42:113-8.

36. Gibbs-Strauss SL, Nasr KA, Fish KM, Khullar 0, Ashitate Y, Siclovan TM, Johnson BF, Barnhardt NE, Tan Hehir CA and Frangioni JV. Nerve-highlighting fluorescent contrast agents for image-guided surgery. Mol Imaging. 2011;10:91-101.

37. Massaad CA, Zhang G, Pillai L, Azhdarinia A, Liu W and Sheikh KA. Fluorescently-tagged anti ganglioside antibody selectively identifies peripheral nerve in living animals. Sci Rep. 2015;5:15766.

38. Gibbs SL, Xie Y, Goodwill HL, Nasr KA, Ashitate Y, Madigan VJ, Siclovan TM, Zavodszky M, Tan Hehir CA and Frangioni JV. Structure-activity relationship of nerve-highlighting fluorophores. PLoS One. 2013;8:e73493.

39. Cotero VE, Kimm SY, Siclovan TM, Zhang R, Kim EM, Matsumoto K, Gondo T, Scardino PT, Yazdanfar S, Laudone VP and Tan Hehir CA. Improved Intraoperative Visualization of Nerves through a Myelin-Binding Fluorophore and Dual-Mode Laparoscopic Imaging. PLoS One. 2015;10:e0130276.

40. Cotero VE, Siclovan T, Zhang R, Carter RL, Bajaj A, LaPlante NE, Kim E, Gray D, Staudinger VP, Yazdanfar S and Tan Hehir CA. Intraoperative fluorescence imaging of peripheral and central nerves through a myelin-selective contrast agent. Mol maging Biol. 2012;14:708-17.

41. Gray D, Kim E, Cotero V, Staudinger P, Yazdanfar S and Hehir CT. Compact Fluorescence and White Light Imaging System for Intraoperative Visualization of Nerves. Proc SPIE /nt Soc Opt Eng. 2012;8207.

42. Hackman KM, Doddapaneni BS, Barth CW, Wierzbicki IH, Alani AW and Gibbs SL. Polymeric Micelles as Carriers for Nerve-Highlighting Fluorescent Probe Delivery. Mol Pharm. 2015;12:4386-94.

43. Wang C, Wu C, Popescu DC, Zhu J, Macklin WB, Miller RH and Wang Y. Longitudinal near infrared imaging of myelination. J Neurosci. 2011;31:2382-90.

EXAMPLE 3: NERVE-TARGETED PROBES FOR FLUORESCENCE-GUIDED INTRAOPERATIVE IMAGING

Abstract

[00395] A fundamental goal of many surgeries is nerve preservation, as inadvertent injury can lead to patient morbidity including numbness, pain, localized paralysis and incontinence. Nerve

identification during surgery currently relies on multiple parameters including anatomy, texture, color and relationship to surrounding structures to distinguish nerves from non-nerve tissues. Using

white light illumination, which is the standard in current operating rooms, the visual difference

between nerves and adjacent tissue can be imperceptible. A nerve-targeted probe, FAM-NP41, that

binds to and highlights rodent motor and sensory nerves following systemic administration was

previously developed. Here it is demonstrated that FAM-NP41 can highlight autonomic nerves

within the prostate gland in living mice and rats with significant nerve to non-nerve contrast in

nerves as small as 50 pm in diameter.

[00396] To translate this methodology for potential clinical use in human patients, phage display was used to identify a novel peptide (HNP401) that selectively binds to human nerves. FAM

HNP401 can bind and highlight multiple human peripheral nerves including lower leg sural nerve,

upper arm medial antebrachial nerve and autonomic nerves including cavernosal nerve surrounding

human prostate gland. The binding domain of HNP401 was identified by sequential deletion of

amino acids from the full-length peptide. HNP401 or an optimized variant could be translated for use

in a clinical setting for intraoperative identification of human nerves to improve visualization and

potentially decrease the incidence of intra-surgical nerve injury.

Introduction

[00397] A fundamental goal of surgery is preservation of nerve function to minimize patient morbidity. Current nerve identification during surgery utilizes non-quantifiable criteria such as

anatomy, texture, color and relationship to surrounding structures. In instances of trauma, tumor

invasion or infection, nerve identification using the above criteria can be even more challenging.

Using white light reflectance, the visual difference between nerves, especially small nerves like the

autonomic nerves within the prostate, and adjacent tissue can be imperceptible. Inadvertent injury

to these thin or buried nerves is one of the most morbid but unintended consequence of surgery which can lead to loss of function, numbness, and surgery induced neuropathic pain [1]. For example, radical prostatectomy (RP) can be performed for localized prostate cancer with excellent locoregional control [2,3] However, even with nerve preserving radical prostatectomy there is a significant risk of erectile dysfunction and/or urinary incontinence, due to inadvertent injury to autonomic nerves or the autonomic neurovascular bundles [4,5]. Preservation of the autonomic neurovascular bundles along the posterolateral aspect of the prostate is an important aspect for functional preservation during RP. The autonomic nerve fibers themselves are rarely visualized, but rather their position is presumed to track along vascular structures. The exact position and distribution of these autonomic nerves are variable from patient to patient complicating the use of anatomical location as the sole method of avoidance [6-8] and injury can occur even in the most experienced hands. Recent studies showed that only 7% of RP patients regained pre-surgical state of full erectile function in the first year [9] and 16% regained baseline erectile function 2 years after prostatectomy [10].

[00398] Tools to improve visualization of the neural structures in the prostate have great potential for reducing morbidity from the radical prostatectomy, as well as applications in many

other nerve preserving surgeries including cancer resection, trauma and reconstructive procedures.

Systemic administration of a nerve imaging agent could allow the labeling of all relevant nerves with

a single probe administration. Previously, reported methods rely on retrograde or anterograde

tracing of individually axonal tracts by direct application of fluorescent dyes to the innervation site

[11, 12]. Styryl pyridinium dyes [13-15], aminostyryl dyes [16-18], oxazine 4 [19, 20], and anti

ganglioside antibodies [21] have been investigated in various preclinical models to detect motor, sensory and autonomic nerves.

[00399] A peptide sequence, Nerve Peptide 41 (NP41), was previously identified through phage display that preferentially binds and highlights peripheral nerve tissue, enhancing visualization

of motor and sensory nerves in live mice after systemic injection [22-24]. This peptide has relatively

low affinity for nerve and rapid blood clearance (compared to antibodies) so it can be visualized a

few hours after systemic injection with almost completely wash out by 24 hours [22]. NP41 has also

been shown to highlight degenerate nerves through the binding to structural laminins in nerve fibers

[24, 25]. We have now used this peptide for intraoperative identification of autonomic nerves in the

prostate of both mice and rats. To allow clinical translation of nerve visualization methods for use in

surgeries involving human patients, we have now used phage display to identify a novel peptide

HNP401 that, when labeled with fluorophore, selectively binds and highlights human nerves.

Fluorescently labeled HNP401 can bind to and highlight human sensory and motor nerves such as

sural, medial antebrachial cutaneous, laryngeal, ansa cervicalis, great auricular nerve and autonomic

nerves like those within and around the prostate gland.

Results

[00400] To visualize the autonomic nerves within the prostate of mice, NP41 peptide conjugated to fluorescein (FAM-NP41) was injected intravenously followed by imaging of prostate

and surrounding tissue after surgical resection. Strong fluorescence from dye that rapidly

accumulates in the bladder hindered visualization of the nerves within the prostate. To enhance

visualization the bladder was surgically drained of urine and sutured prior to imaging. The urethra,

an anatomically distinct structure, is never emptied of urine as the mice are alive for the duration of

the experiment, resulting in a continuous passage of urine carrying metabolized peptide-dye to the

bladder via the urethra. To aid future research, we have demonstrated using a fluorescent

quenching dye (both directly injected in the bladder and through oral administration) to reduce high

bladder fluorescence, as an alternative to surgical draining of the bladder (Figure 21). FAM-NP41 was

injected at doses ranging from 150-600 nmoles (~16 to 66mg/kg) with a 600 nmoles (~30nmol/g)

dose showing optimal autonomic nerve contrast (Figures 16A-16G). Low magnification fluorescent

image show highlighting of a single nerve fiber running adjacent to the urethra (Figure 16A). The

nerve is extremely faint in a high magnification image using white light reflectance (Figure 16B) but becomes distinctly visible with FAM-NP41labeling (Figure 16C). To quantify nerve detection a total

of 10 mice were injected with 600nmoles FAM-NP41 and signal intensity was measured for nerve

versus adjacent non-nerve tissue using both fluorescence and white light reflectance. Values to the

right of the line indicate that there is improved visualization with fluorescence compared to

reflected light. Average nerve to non-nerve signal intensity with fluorescent guidance was 1.256

0.14 (n=12, p<0.001) compared to 1.086 ±0.07 (n= 12) for white-light reflectance (Figure 16D).

[00401] Because prostate nerves in mice were very small and challenging to image (i.e. requiring high dose of FAM-NP41) we extended our study to the visualization of autonomic nerve

within the prostate of rats. To visualize autonomic nerves in male Sprague Dawley rats, FAM-NP41

was injected intravenously at a dose of 12nmol/gram, followed by imaging. This is a 2.5X lower dose

relative to weight compared to the 600nmols used in 20 gram mice. Useful labeling occurred 2 to 6

hours after intravenous administration which was visualized using a customized fluorescence

dissecting microscope. FAM-NP41 nerve highlighting enables visualization of nerve fibers running

through the middle of the rat prostate (Figure 16E). Higher magnification imaging showed that FAM

NP41 additionally highlighted autonomic nerve branches surrounding the neurovascular bundle

(Figure 16G) which travel within the fatty capsule of the prostate gland. These branching nerves

were not visible using white light reflectance imaging (Figure 16F). To quantitate selective labeling of autonomic nerves in rats, nerves within the prostate gland were imaged with both fluorescence and

white light reflectance. Average nerve to non-nerve signal intensity from fluorescence was 1.275±

0.02 (n=3) compared to 1.083 ±0.01 (n=3) for white light reflectance. To show applicability to intra

surgical imaging we show that similar nerve contrast was observed in live rats using a clinical grade

Zeiss Pentero imaging system (Figure 16H). The Zeiss Pentero scope which is approved for clinical

use overlays the fluorescent image from FAM-NP41 (yellow) on the white light image with data

collection in real time (Figure 16H). Recordings during surgical manipulation show fluorescent fibers

within the prostate that clearly present as nerves that are detectable using NP41-FAM fluorescent

guidance (data not shown). To confirm that fluorescently labelled structures were indeed nerves,

fluorescent surgical guidance was used in real time to selectively dissect out fluorescent fibers that

were thought to be nerves (Figure 22: A). Dissected fluorescent fibers were then positioned

vertically and flash frozen in OCT embedding compound. Vertical cross sections were imaged using

fluorescence to show that suspected nerve fibers were centered on slides (Figure 22: B). Fibers were confirmed to be nerve as they were fluorescently labelled using dual immunohistochemically

analysis with antibodies against either fluorophore (Figure 22: C) or tyrosine hydroxylase (Figure 22:

D), a known marker for unmyelinated autonomic nerves. No immunostaining was detected in the

absence of primary antibody (Figure 22: E)

[00402] To enable translation of a nerve-illumination peptide for use in human patients, phage display was performed to identify human nerve binding peptides using an m13 phage library

expressing 16 random amino acid sequences on the N-terminus of gill (Creative Biolabs). Phage were

selected using iterative rounds of selection for binding to human sural nerve with negative selection

to muscle and fat. Counter selection to muscles and fat was done by pre-absorbing library with these

tissues prior to selection for binding to human nerve. Individual phage were sequenced after each

round of selection and three specific sequences SGQVPWEEPYYVVKKS (HNP401; SEQ ID NO:1), and

WEYHYVDLNWTSQHPQ (HNP402; SEQ ID NO:2) DLPDIIWDFNWETAG (HNP403; SEQ ID NO:3) were

highly enriched after 5 and 6 rounds.

[00403] To test the affinity of selected phage display peptides for binding to human nerves, they were chemically synthesized by solid-phase synthesis and labeled with fluorescein at the C

terminus. Peptides were topically applied to sections of surgically harvested human sural nerve and temporalis muscle to determine nerve to muscle contrast for selected peptides and controls (Figure

23). Controls including free dye (carboxyfluorescein) were also tested on various nerves from

multiple patient tissues to confirm specificity of peptide dye conjugates for binding human nerve (Figure 24). Free non-reactive dye control, such as carboxyfluorescein, was shown to have only weak

non-specific binding and are not efficacious for topical applications. FAM-HNP401 yielded the

highest contrast and was shown to be superior to the previously reported rodent nerve binding

peptide FAM-NP41 [22], when topically applied to human sural nerve (Figure 17). To quantify

differential binding to nerve versus muscle, fluorescence signal intensity was measured for ROls

from the perineurium of select nerves and human temporalis muscle. FAM-HNP401 showed

selective binding to human sural nerve with 10.9X fluorescent signal intensity (1374.44 425.96)

compared to FAM-NP41 (126.17 ± 61.03) (Figure 17D, p=0.009, Student's t-test, unpaired). Nerve to

muscle contrast was comparable at 3.03 0.57 for FAM-HNP401 and 2.28 i 0.96 for FAM-NP41

(Figure 17H, p=0.236, Student's t-test, unpaired).

[00404] FAM-HNP401 was also tested topically on ex-vivo tissue for labeling of mouse facial nerve with surrounding muscle where it did not perform as well as FAM-NP41 (Figure 25: J-M). For

comparison, ex-vivo tissue labeling of human laryngeal nerve with surrounding muscle with FAM NP41 and FAM-HNP401 is shown (Figures 25: F-1). Autofluorescence of human nerve without

treatment of peptide dye conjugate was negligible compared to signal intensity acquired after

topical application FAM-HNP401 (Figure 26). FAM-HNP401 also has a 2.3X higher signal intensity for

in-vivo binding to mouse sciatic nerve compared to FAM-NP41(Figure 18 A-C). Nerve to surrounding

muscle contrast is comparable for the two peptides (Figure 18D). FAM-HNP401 also highlighted rat

sciatic nerve (Figure 18E) and prostate nerve (Figure 18F) at a dose of 2Imole (~54mg/kg) when

imaged 3 hours post injection. The bladder was drained with a syringe and sutured to avoid spillage

and contamination around prostate. The collected urine was analyzed by mass spectrometry and as

expected fragments of the peptide with dye attached were detected indicating peptide in bladder

was partially metabolized (Figure 27). Autonomic nerves within the prostate and adjacent to the

vascular bundle can be easily visualized when imaged at higher magnification using a dose of

0.5.tmole (13.4mg/kg) FAM-HNP40110 mins after probe injection (Figure 18G). Blood clearance of

FAM-HNP401 showed a half-life of 30 minutes which is similar to FAM-NP41 (Figure 18H). Optimal

nerve contrast was detected using 50-100M (Figures 25: A-E) with low concentration (10 M) high

resolution confocal imaging showing that FAM-HNP401 binds with higher affinity to perineurium,

epineurium and endoneurium while being excluded from axons (Figure 25: N). FAM-HNP401 signal

from human nerve saturates by 100iM while the signal from FAM-NP41 continues to increase even at 375IM but the signal intensity remains much lower than that of HNP401 applied at the same concentration (Figures 25: F and H). Stability of FAM-HNP401 in human plasma at 5 minutes and 2 hours was determined by incubation of peptide dye conjugate in human serum prior to analysis by mass spectrometry. For analysis the area under the curve at 450nm and the corresponding mass of

FAM-HNP401 was determined after injection of a fixed volume of analyte into the LC-MS (Figures

28:A-B). For comparison, we also tested the stability of FAM-NP41 in human plasma (Figures 28:C

D). Integration of the peak area at 5 minutes and 2 hours indicates that both FAM-HNP401 and FAM

NP41 were stable in human serum. Area of extracted ion-current was used to determine peptide

quantitation. No degradation of peptide-FAM conjugate was observed, with identical concentration

detected at 5min and 2hours of incubation with human plasma from analysis of the ion current. Peptides were analogously tested and shown to be stable in rat cerebrospinal fluid following 2 hour

exposure (Figures 28:E-F).

[00405] FAM-HNP401 and FAM-NP41 were tested for binding to autonomic nerves, isolated from the prostate glands of two human patients (Figures 19 and 29). FAM-HNP401 (Figures 19A and

29A) showed a significantly higher fluorescent signal in autonomic (cavernosal) nerves compared to FAM-NP41 (Figures 19B and 29B). Quantitation was not done because only 2 patient samples were

available for testing as nerve resection during radical prostatectomy is only performed in instances

of gross capsular invasion. Labelled fibers were confirmed as nerve using anti-neurofilament

antibody SM1312 (red) with DAPI (blue) to show nuclear labeling (Figure 19C). H&E staining also

confirmed label tissue as nerve by histology (Figure 19D). SM1312 does not stain perineurium due to

the lack of neurofilament fibers in this region of the nerve bundle. SM1312 staining shows that the

tissue isolated is nerve due to staining of neurofilament structures that support the axons. Similar

staining using FAM-HNP401 was obtained for another sensory nerve (anti-brachial cutaneous)

isolated from human arm showing the broad nerve binding activity of HNP401 (Figures 19E-H).

[00406] To optimize and attempt to determine the core binding domain of HNP401, systematic deletion of two amino acids from the C or N terminus was performed (see, for example,

Figure 30) followed by binding analysis on human sural nerve sections (Figure 20). In each case

nerve binding and signal intensity was normalized to the parent FAM-HNP401 peptide (Figure 20J).

Removal of the C-terminal serine (C-2) was tolerated but upon removal of lysine (C-4) the solubility

and binding was reduced dramatically with a normalized average signal intensity of 0.49 ±0.11for

nerve binding of HNP401-C-4 (Figure 20F). Deletion of amino acids from N terminal is mostly well

tolerated. Removal of the N-terminal serine and glycine improved nerve selective binding about 2 fold with a normalized average signal intensity of 2.02 0.65 for HNP401-N-2 (Figures 20A and 20J, p=0.026, Student's t-test, unpaired, one-tail). HNP401-N-4 has non-polar amino acids on its N terminus which reduced binding to a normalized average signal intensity of 0.56± 0.18 (Figure 20B).

Removal of non-polar amino acids, tryptophan and proline, restored some binding intensity back to levels of FAM-HNP401 with HNP401-N-6 (Figure 20C) and HNP401-N-8 (Figure 20D) having

normalized average nerve signal intensities of 1.0 0.34 and 0.98 0.31. The restored binding

efficiency may be due to improved solubility minimizing micro-aggregation that occurred when very

hydrophobic residues are present at the N terminus of peptide. C and N terminal deletion studies of

HNP-401 indicate the core binding domain likely includes PYYVVKK with the N-terminal residues

QVPWEE contributing to enhanced binding detected with HNP401-N-2. Normalized nerve to

temporalis muscle contrast for HNP401-N-2 gave a 3-fold increase with respect to FAM-HNP401 (Figure 20K, p=0.011, Student's t-test, unpaired, one-tail).

Discussion

[00407] Various tracer substances have long been used to map the connectivity in the nervous system although most of them have depended on anterograde or retrograde tracing after

local application [11, 12, 26, 27]. Transport of tracers is relatively slow with contrast developing as

dye moves away from a the injection site [26, 27]. It is likely impractical to label the large areas

exposed for surgeries by using these methods as multiple nerve tracts would have to be identified

and independently labelled. There are reports of tracking retrograde neurovascular bundle and

major pelvic ganglion with lipophilic dyes in rodents [4, 28]. More recently, styryl pyridiniurn dyes

[13-15,aminostyryl dyes[16-18], oxazine 4 [19, 20], and anti-ganglioside antibodies [21] have been

investigated in various preclinical models to detect motor, sensory and autonomic nerves. Dyes

alone have no selective mechanism for nerve targeting but typically accumulate in the myelin.

Myelin is known to be present in low abundance or be absent in autonomic nerves which could limit

the use of free dyes to highlight these fine but crucial nerves [29, 30]. Topical and epidural

application of free dyes has been used to locally label nerves in animal models however these approaches may be limited in flexibility during human surgeries as tissue is removed and the field of

view changes [20, 31]. Anti-ganglioside antibodies have specific targeting but have long blood half

lives which would likely require injection multiple days before surgery and may be more likely to

elicit an immune response[32, 33]. Systemic injection of fluorescently labeled peptides to label

nerves overcomes the major disadvantages of these tracers by labeling all nerves in the body with a

single injection of peptide dye conjugate. We previously reported on NP41 for binding rodent motor

and sensory nerves and now demonstrate its potential application to the identification of fine autonomic nerves in rodent models. We found an average increase of 17% in nerve to non-nerve signal using fluorescence imaging compared to contrast obtained by to white light reflectance. This is a significant accomplishment given the unmyelinated nature of these nerves and their ultra-fine structure. However, topical application of NP41 to human ex-vivo provided little contrast compared to muscle. To enhance highlighting of human nerves with have now identified HNP401, a novel peptide that binds to and highlights human motor/sensory and autonomic nerves.

[00408] We expect the FAM-HNP-401 or optimized analog could enable clinical translation of nerve visualization methods for use in surgeries involving human patients. Fluorescently labeled HNP401 can bind and highlight human sural, medial antebrachial cutaneous, laryngeal and

autonomic nerves within and around the prostate gland. FAM-HNP401 show high signal intensity

and reproducible labelling of nerve bundles compared to its dye control of carboxyfluorescein.

Carboxyfluorescein shows low signal and non-specific binding to nerve on topical human nerve

sections. Dyes such as FITC-isothiocyanate cannot be used as the control as they will react with all

nucleophilic side chains of proteins exposed by cross-sectioning in unfixed tissue. Additionally, FITC

dextran, although clinical used, is not a viable control for our experiments as it labels vasculature

including micro blood vessels deep within the nerve cross section and is a marker for nerve injury

and neuropathic pain [34]. In addition, its large size affects pharmacokinetic profile of the dye. FAM

HNP401 consistently gave 10-fold higher signal for binding human nerve compared to our previously

identified FAM-NP41 peptide dye conjugate. Higher signal intensity is an advantage for real-time

imaging requiring short exposure times. HNP401 also showed a 3-fold contrast for nerve to muscle

on topical sections in human ex-vivo tissue. FAM-HNP401 has a blood clearance profile similar to NP41 in mice [22]. FAM-HNP401 binds to myelinated and unmyelinated nerves. SM1312 antibody,

which labels neurofilament does not colocalized with FAM-HNP401 staining demonstrating that

FAM-HMP401 does not bind axons, but preferentially binds the perineurium, and therefore may be

less likely to affect nerve conductivity. It is this staining pattern that leads us to believe HNP401-FAM

is binding structural protein(s) in the perineurium. Polar amino acids at the C terminus appear to be

needed for both solubility and binding as removal either caused peptide to become significantly less

soluble or show decreased binding affinity to nerves. Removal of 2 amino acids on the N terminus

increased nerve binding but further deletions negatively affected both solubility and binding.

Attaching solubilizing groups like short PEGs may restore binding to truncated variants.

[00409] For our initial studies nerve highlighting peptides HNP401 has been coupled to fairly short wavelength fluorescein derivative to make it compatible with dual nerve/tumor imaging with

Cy5/Cy7 ratiometric activatable cell penetrating peptides that are currently in phase 11 clinical testing

for detection of cancer (NCT03113825). Longer wavelength IR or near IR dyes such as indocyanine

green (ICG), IRdye800 would potentially allow nerves to be imaged deeper below the surface in surgically exposed tissue after attachment to HNP-401. Free oxazine 4 has also been recently used to

highlight nerves in preclinical models and targeting could be enhanced by coupling to targeting

peptides like HNP401. Although our preferred method of application is systemic, topical application

is an option with some procedures. Such topical application of dye to the exposed surface followed

by a washing to remove unbound dye has been used to image nerve in animal models [20]. Dyes

such as 4-di-2-asp have also been used for topically application to nerves but it has the disadvantage

of being toxic to nerves due its binding to mitochondria in nerve terminals [35]. Antibodies can be

applied intravenously or topically and have some advantages including high affinity and a defined

binding target, however as reported with the anti-ganglioside antibody they require long circulation

times for accumulation and washout to develop optimal nerve contrast.

[00410] In in-vivo rodent studies, we found that peripheral motor and sensory nerve can be labeled in mice at a dose of 150 nmoles FAM-NP41 which would easily scale to human dosing [36,

37]. Autonomic nerve labeling required a significantly higher dose in mice (600nmols) so higher

affinity peptides like HNP401 or improved variants may be required for advancement to clinical

dosing. Interestingly, although higher dosing was required to visualize very small autonomic nerve in

rodents (as small as 50 m) labeling of significantly larger human prostate nerves (~750 m) may be

accomplished at a significantly reduced dose. Consistent with the conclusion that larger nerve can be

highlighted with a lower dose, we were able to visualize nerve in rat prostate with a 40% dose NP41.

Neither NP41 nor HNP401 permanently or covalently bind to nerve bundles as they both washes out

with little remaining signal after 24 hours. Structural proteins including laminins 421, 211 have been

identified as the binding targets for NP41 [25]. While the binding targets for HNP401 is yet to be

determined imaging data shows non-axonal binding pattern similar to NP41 indicating it may also

bind structural nerve proteins. One significant characteristic of HNP401 compared to lipophilic dyes

is that it does not require the presence of myelin and we have shown that it can bind and highlight

the neurovascular bundle as well as the cavernosal nerve within the prostate. These nerves are important in urological applications and do not have high levels of myelination [29, 30]. We

anticipate that preservation of nerves in this context represent one of the most urgent unmet

clinical needs [38] for nerve imaging technology. The ability of FAM-HNP401 to highlight these

nerves represents a significant advantage over competing nerve binding agents that are selective for

myelin [39] and incorporate into axons [21].

Methods

Probe synthesis

[00411] FAM-NP41 was synthesized as previously described [22]. A Prelude peptide synthesizer and standard Fmoc solid phase peptide synthesis was used to generate peptides with

sequence acetyl-SGQVPWEEPYYVVKKSSGGC-CONH 2 [HNP401], acetyl-WEYHYVDLNWTSQHPQGGC

CONH 2 [HNP402], acetyl-DLPDIIWDFNWETAGGC-CONH 2 [HNP403], each peptide having a C-terminal

"GGC" linker. Carboxyfluorescein was conjugated to the C-terminal cysteine using 5-fluorescein

maleimide [Anaspec] in the presence of N-methylmorpholine in DMSO. Peptides were purified on

Agilent LCMS using a Phenomenex 5um C18 Luna with mass and purity > 95% confirmed by LC-MS.

Truncated HNP401 peptides as listed in (see, for example, Figure 30) were synthesized and purified

using the same configuration and method described above.

Animals

[00412] Wild-type male SKH1 mice (Charles River, Wilmington, MA) weighing 20-30 grams were used for testing of peptide dye conjugates. Male Sprague-Dawley rats weighing 100 to 250

grams were used for in-vivo testing of dye conjugates with dose being adjusted for based on animal

size. Protocols for use of animal were approved by the Institutional Animal Care and Use Committee

at University of California San Diego (Protocol number S05536).

In vivo imaging

[00413] Following anesthesia with intra-peritoneal injection of ketamine (80mg/kg) and midazolam (40mg/kg), FAM-NP41 or its variants were administered into mice retro-orbitally. After a

washout period of 2-4 hours, the animals were anesthetized with ketamine (50 mg/ml) and xylazine

(20mg/ml). The bladder and prostate were exposed through a midline abdominal incision. The

autonomic nerve along the cavernosal vessel in the prostate was imaged and recorded using a

custom-made surgical imaging system. This system is a modified from Olympus MVX10 scope

capable of hi-resolution fluorescence, RGB reflectance and realtime overlay with zoom from 0.6 to

5.7 cm field of view. ImageJ was used for quantitative analysis of nerve contrast for each peptide dye

conjugate tested. Images of autonomic nerve in prostate were selected from the recorded files and

magnified 300-400% prior to selection of ROI and measurement. Nerves and adjacent non-nerve

tissues ROls were hand-selected using polygonal selection tool at the same location from both of

reflectance and fluorescence images. The mean and standard deviation of the pixel intensities within

the selected areas were compared for nerves (mean = I, SD = o,) and adjacent background tissue

(mean=l, SD=). Nerve to non-nerve contrast was calculated after background subtraction with formula | In-I bI / (o2 + n 2 0.. For imaging the nerves in the prostate gland of male rats peptide

dye conjugate were injected retro-orbitally. FAM-HNP401 was injected at a concentration of 13mgs/kg followed by imaging after 15 minutes or alternatively a dose of 52mgs/kg was used with imaging after 3 hours. Live animal surgery was performed under a ketamine-xylazine cocktail according to IACUC protocol. Sterile technique was used to expose the prostate; bladder was drained with a small syringe and sutured. The surgical field was washed with sterile saline prior to imaging. Mann-Whitney test was used to analyze data for both mice and rats to compare nerve intensity and nerve to non-nerve contrast between white light reflectance and fluorescence.

Confocal Imaging parameters

[00414] Confocal data for Figure 17 was acquired with 488nm laser line, 101m sections on glass at loX magnification, 0.45 NA air objective lens. Gain set to 50, power set to 0.5% of laser power, pixel dwell value of 1.2 ls, aperture size of 1.2 am and a pixel size of 0.26 with a 2k by 2k size image. We used the Nyquist feature and acquired images as tiles to get maximum resolution.

[00415] Data for Figures 19A and 19B was acquired with 488nm laser line, 101m sections on glass at lOX magnification, 0.45 NA air objective lens. Gain set to 40, power set to 3% of laser power, pixel dwell value of 1.2 ls, aperture size of 1.2 pm and a pixel size of 0.26 am/px with a 2k by 2k size image.

[00416] Data for Figures 19E and 19F was acquired with 488nm laser line, 101m sections on glass at lOX magnification, 0.45 NA air objective lens. Gain set to 40, power set to 1% of laser power, pixel dwell value of 1.2 ps, aperture size of 1.2 pm and a pixel size of 0.3am/px with a 2k by 2k size image.

[00417] SM1312 neurofilament antibody and Dapi staining were imaged at OX magnification, 0.45 NA air objective lens, NA with gain of 50, power set to 5% of laser power for 405nm laser line and gain of 100, power set to 50% of laser power for 640nm laser line. We used a pixel dwell of 3.2 ps, aperture size of 1.2 pm and image size of 2k by 2k per tile resulting in a pixel size of 0.29 lm/px.

[00418] Data for Figure 26 was acquired with 488nm laser line, 10pm sections on glass at ioX magnification, 0.45 NA air objective lens. Gain set to 40, power set to 3% of laser power, pixel dwell value of 2.4 as, aperture size of 1.2 am and a pixel size of 0.3am/px with a 2k by 2k size image.

[00419] Dose response data set of FAM-HNP401 on human nerve tissue (Figures 25:A-E) was acquired with 488nm laser line, 10pm sections on glass at oX magnification, 0.45 NA air

objective lens. Gain set to 40, power set to 3% of laser power, pixel dwell value of 1.2as, aperture

size of 1.1 pm and a pixel size of 0.3am/px with a 2k by 2k size image.

[00420] Data for Figure 29 was acquired with 488nm laser line, 10am sections on glass at 25X magnification, 1.10 NA water immersion lens. Gain set to 40, power set to 3% of laser power,

pixel dwell value of 2.2 ps, aperture size of 1.2 pm and a pixel size of 0.11 am/px with a 2k by 2k size

tiled image.

Phage display

[00421] Phage display was done using a custom synthesized m13 phage libraries (diversity ~109) expressing 16 random amino acid on the N-terminus of gill (Creative Biolabs). The phage

library was processed through selections for binding to freshly resected or frozen human nerves as

similarly describe for the identification of NP41 which bound mouse nerves [22]. Library was processed through up to 6 binding and wash cycles. Prior to positive selection phage were counter

selected for high affinity muscle and fat tissue binder by pre-absorbing library with these tissues. For

positive selection phage libraries were mixed directly with human sural nerve tissue and incubated

for up to 2 hours at 4 °C. Following incubation, tissue phage mixtures were centrifuged and washed

with PBS. Tissue pellets with bound phage were then homogenized, mixed with TG1 bacteria and

plated on LB agar plates. Colonies were counted to determine titer followed by selecting single

colonies for DNA preparation and sequencing. After each round of selection phage were pooled and

amplified for iterative selection. Phage that were bound at each round were sequenced and repeats

noted. Duplicate phage as shown in results were identified after 5 and 6 rounds of selection.

Topical application on tissue sections and imaging

[00422] Human sural nerve, antebrachial nerve and laryngeal nerve and temporalis muscle were obtained under IRB protocol number 130837 for Dr. Quyen Nguyen. Human peripheral nerves

(typically sural) were obtained from patients undergoing nerve resection procedures. Human nerves

from prostate gland of two patients were acquired under Moores Cancer Centre Biorepository IRB

protocol number 090401. Tissue were sectioned and mounted on glass slides or Cryojane tape.

Tissue sections were placed in a humidifier chamber for 30 min before application of the peptide

solution. Peptides were diluted to appropriate concentration in .5X HBSS prior to topical application.

50.l of peptide solution of with known concentration (1pM to 375pM) were applied to 10pm nerve sections on tape or slides and incubated for 30 minutes in a humidifier chamber. After incubation

with peptide nerve sections were washed with twice with 0.5X HBSS and once with IX PBS. A cover

slip was applied and slides were imaged immediately on either a Zeiss Lumar dissecting scope of

Nikon Al confocal microscope. For confocal imaging tissue sections of 10am thickness were imaged

with 488nm laser excitation 515 (25) and a 10X air objective and a 0.26Mm/pixel size. For

immunohistochemistry the confocal Images were acquired with a 20X air objective at 0.4am/pixel.

Image analysis

[00423] Image J was used to analyze and compare images acquired using the confocal microscope and the Lumar dissecting scope. For each experimental set where probes were

compared, we kept the acquisition parameters identical so as to directly compare the data obtained.

During the experiment, it is clear that FAM-HNP401 had the brightest signal in our topical application

experiments. All raw image files for a given experimental cohort were loaded at the same time into

Image J as 16-bit tiff images. We then levelled the image for tissue treated with FAM-HNP401. Once

these levels were set, the settings are propagated to all images in one step using Image J. The

brightest image is set as the benchmark for all other images in the cohort to avoid saturating when the leveling is propagated. For quantifying the images, regions of interest (ROI) were drawn and the

signal counts measured in image J. For Figure 5, even though FAM-HNP401-N-2 is the brightest, for

consistency we choose FAM-HNP401 to level and normalize signal counts.

Immunofluorescence of autonomic nerves from rat prostate

[00424] Suspect unmyelinated nerve tissue was taken from prostate gland of male rat after in-vivo intravenous injection of TAMRA-NP41 (0.5 moles or 11.3mg/kg for 100gm rat) visualized on

custom-made surgical fluorescence imaging system based on an Olympus dissecting microscope.

5im cryosections of the tissue were generated using a Leica Cryostat and mounted on Cryojane

tape. Tissue sections were fixed for 10 min with 4% para-formaldehyde in 1X PBS followed by a 1X

PBS rinse. A 1:2000 dilution of monoclonal antibody against TAMRA [Thermofisher Scientific Cat. No.

MA1-041] (or polyclonal antibody against tyrosine hydroxylase [Cell Signaling Technologies Prod. No.

2792S]) in 10 % goat serum in PBS were applied; 20 I per section and incubated overnight at room

temperature followed by a 1X PBS wash. A 1:500 dilution of biotinylated anti-mouse secondary

antibody was applied in 10% goat serum in PBS to sections for 2 hours followed by a 1X PBS wash.

Vector RTU (avidin biotin complex) or Alexa 405 streptavidin was applied for 1 hour followed by a 1X

PBS wash. Tissue was wet-mounted on slides with 1X PBS. Confocal Image was acquired with 20 x air

objective at resolution of 0.4 m/pixel.

Immunofluorescence for neurofilament

[00425] Fresh viable human nerve tissue was obtained from prostatectomy and frozen in OCT blocks. 10im cryosections of tissue were mounted on glass True Bond slide. Hydrophobic

barrier pen was applied to the glass around each section. Tissue sections were fixed using 2%

paraformaldehyde prepared in IX PBS and washed 4 times with 1X PBS. 100d of blocking buffer

(0.01% Triton X solution, 1%BSA in 10% normal goat serum [Life technologies 50062Z]) was applied

for 30min to each tissue section. The tissue was then washed 4 times with IX PBS and a 1:1000

dilution of neurofilament antibody SM1312 antibody [Biolegend Cat. No. 837904] was applied to the

tissue for overnight incubation at 4°C. Tissue was washed 6 times with PBST. A 1:1000 dilution of

anti-mouse secondary antibody Alexafluor 555 was applied to the sections for 2 hours at 4°C

followed by washing with 1X PBS. Prolong Gold Anti-fade reagent with DAPI [Life Technologies

P36931] was added prior to cover slipping and imaging.

H&E staining protocol

[00426] Tissue sections were fixed for 1 minute in 1:110% buffered formaldehyde and 200 proof ethanol. Slides were then washed with water and immersed in hematoxylin stain for 2

minutes. Slides were then washed with distilled water and immersed in bluing solution for 30

seconds. Slides were washed with distilled water and immersed in eosin solution for minute

followed by wash with distilled water. Slides were sequentially dipped in 50%, 95% and 100%

ethanol to remove water. Slides were air dried and dipped in citrisolv before mounting a cover-slip

with non-xylene mounting solution and imaged on the Hamamatsu Nanozoomer using bright-field at

20X magnification.

Blood clearance for HNP401-FAM

[00427] Five 8-week-old SKH male mice were injected intravenously with 100nmol

[10.75mg/kg for 25gm mouse] of FAM-HNP401 in 100 of sterile water. Prior to blood draw mice

were anesthetized with a 1:1 cocktail of ketamine: midazolam. Tail pricks were performed at1min,

10min, 20min, 30min, 1 hr, and 2 hrs after injection to collect 5 Iwhole blood which was dissolved

in 100 Agilent ICP-MS tuning buffer. Samples were centrifuged and equal volume of supernatants

were analyzed using a Tecan fluorescence plate reader.

References

[00428] [1] Borsook D, Kussman BD, George E, Becerra LR, Burke DW. Surgically induced neuropathic pain: understanding the perioperative process. Annals of surgery. 2013;257:403-12.

[00429] [2] D'Amico AV, Whittington R, Malkowicz SB, Schultz D, Blank K, Broderick GA, et al. Biochemical outcome after radical prostatectomy, external beam radiation therapy, or interstitial radiation therapy for clinically localized prostate cancer. Jama. 1998;280:969-74.

[00430] [3] Walsh PC. Radical prostatectomy for localized prostate cancer provides durable cancer control with excellent quality of life: a structured debate. The Journal of urology. 2000;163:1802-7.

[00431] [4] Yamashita S, Kato R, Kobayashi K, Hisasue Si, Arai Y, Tsukamoto T. Nerve injury related erectile dysfunction following nerve-sparing radical prostatectomy: A novel experimental dissection model. International journal of urology. 2009;16:905-11.

[00432] [5] Stanford JL, Feng Z, Hamilton AS, Gilliland FD, Stephenson RA, Eley JW, et al. Urinary and sexual function after radical prostatectomy for clinically localized prostate cancer: the Prostate Cancer Outcomes Study. Jama. 2000;283:354-60.

[00433] [6] Walsh PC. Anatomic radical prostatectomy: evolution of the surgical technique. The Journal of urology. 1998;160:2418-24.

[00434] [7] Tewari A, Peabody JO, Fischer M, Sarle R, Vallancien G, Delmas V, et al. An operative and anatomic study to help in nerve sparing during laparoscopic and robotic radical prostatectomy. European urology. 2003;43:444-54.

[00435] [8] Nandipati KC, Raina R, Agarwal A, Zippe CD. Nerve-sparing surgery significantly affects long-term continence after radical prostatectomy. Urology. 2007;70:1127-30.

[00436] [9] Koehler N, Holze S, Gansera L, Rebmann U, Roth S, Scholz HJ, et al. Erectile dysfunction after radical prostatectomy: the impact of nerve-sparing status and surgical approach. International journal of impotence research. 2012;24:155-60.

[00437] [10] Nelson CJ, Scardino PT, Eastham JA, Mulhall JP. Back to Baseline: Erectile Function Recovery after Radical Prostatectomy from the Patients' Perspective. The journal of sexual medicine. 2013.

[00438] [11] O'Malley MR, Wittkopf JE, CutlerJL, Labadie RF, HackettTA, Haynes DS. Fluorescent retrograde axonal tracing of the facial nerve. The Laryngoscope. 2006;116:1792-7.

[00439] [12] Marangos N, Illing R-B, KrOger J, Laszig R. In vivo visualization of the cochlear nerve and nuclei with fluorescent axonal tracers. Hearing research. 2001;162:48-52.

[00440] [13] De Proost I, Pintelon I, Brouns I, Timmermans J-P, Adriaensen D. Selective visualisation of sensory receptors in the smooth muscle layer of ex-vivo airway whole-mounts by styryl pyridinium dyes. Cell and tissue research. 2007;329:421-31.

[00441] [14] Naskar R, Wissing M, Thanos S. Detection of early neuron degeneration and accompanying microglial responses in the retina of a rat model of glaucoma. Investigative ophthalmology &visual science. 2002;43:2962-8.

[00442] [15] Papworth GD, Delaney PM, Bussau U, Vo LT, King RG. In vivo fibre optic confocal imaging of microvasculature and nerves in the rat vas deferens and colon. Journal of anatomy. 1998;192:489-95.

[00443] [16] Gibbs-Strauss SL, Nasr KA, Fish KM, Khullar 0, Ashitate Y, Siclovan TM, et al. Nerve-highlighting fluorescent contrast agents for image-guided surgery. Molecular imaging. 2011;10:7290-2010.

[00444] [17] Cotero VE, Kimm SY, Siclovan TM, Zhang R, Kim EM, Matsumoto K, et al. Improved intraoperative visualization of nerves through a myelin-binding fluorophore and dual mode laparoscopic imaging. PloS one. 2015;10:e0130276.

[00445] [18] Stankoff B, Wang Y, Bottlaender M, Aigrot M-S, Dolle F, Wu C, et al. Imaging of CNS myelin by positron-emission tomography. Proceedings of the National Academy of Sciences. 2006;103:9304-9.

[00446] [19] Park MH, Hyun H, Ashitate Y, Wada H, Park G, Lee JH, et al. Prototype nerve specific near-infrared fluorophores. 2014.

[00447] [20] Barth CW, Gibbs SL. Direct Administration of Nerve-Specific Contrast to Improve Nerve Sparing Radical Prostatectomy. Theranostics. 2017;7:573-93.

[00448] [21] Massaad CA, Zhang G, Pillai L, Azhdarinia A, Liu W, Sheikh KA. Fluorescently tagged anti-ganglioside antibody selectively identifies peripheral nerve in living animals. Scientific reports. 2015;5:15766.

[00449] [22] Whitney MA, Crisp JL, Nguyen LT, Friedman B, Gross LA, Steinbach P, et al. Fluorescent peptides highlight peripheral nerves during surgery in mice. Nature biotechnology. 2011;29:352-6.

[00450] [23] Wu AP, Whitney MA, Crisp JL, Friedman B, Tsien RY, Nguyen QT. Improved facial nerve identification with novel fluorescently labeled probe. The Laryngoscope. 2011;121:805-10.

[00451] [24] Hussain T, Mastrodimos MB, Raju SC, Glasgow HL, Whitney M, Friedman B, et al. Fluorescently labeled peptide increases identification of degenerated facial nerve branches during surgery and improves functional outcome. PloS one. 2015;10:e0119600.

[00452] [25] Glasgow HL, Whitney MA, Gross LA, Friedman B, Adams SR, Crisp JL, et al. Laminin targeting of a peripheral nerve-highlighting peptide enables degenerated nerve visualization. Proceedings of the National Academy of Sciences. 2016;113:12774-9.

[00453] [26] K6bbert C, Apps R, Bechmann I, Lanciego JL, Mey J, Thanos S. Current concepts in neuroanatomical tracing. Progress in neurobiology. 2000;62:327-51.

[00454] [27] Richmond FJR, Gladdy R, Creasy JL, Kitamura S, Smits E, Thomson DB. Efficacy of seven retrograde tracers, compared in multiple-labelling studies of feline motoneurones. Journal of neuroscience methods. 1994;53:35-46.

[00455] [28] Davila HH, Mamcarz M, Nadelhaft I, Salup R, Lockhart J, Carrion RE. Visualization of the neurovascular bundles and major pelvic ganglion with fluorescent tracers after penile injection in the rat. BJU international. 2008;101:1048-51.

[00456] [29] Schaumburg HH, Zotova E, Cannella B, Raine CS, Arezzo J, Tar M, et al. Structural and functional investigations of the murine cavernosal nerve: a model system for serial spatio temporal study of autonomic neuropathy. BJU international. 2007;99:916-24.

[00457] [30] Karam I, Droupy S, Abd-Alsamad I, Korbage A, Uhl J-F, Benoit G, et al. The precise location and nature of the nerves to the male human urethra: histological and immunohistochemical studies with three-dimensional reconstruction. European urology. 2005;48:858-64.

[00458] [31] Liu W, Gu R, Zhu Q, Xiao C, Huang L, Zhuang X, et al. Rapid fluorescence imaging of spinal cord following epidural administration of a nerve-highlighting fluorophore. Theranostics. 2017;7:1863-74.

[00459] [32] Descotes J. Immunotoxicity of monoclonal antibodies. 2 ed: Taylor & Francis. p. 104-11.

[00460] [33] Chames P, Van Regenmortel M, Weiss E, Baty D. Therapeutic antibodies: successes, limitations and hopes for the future. British journal of pharmacology. 2009;157:220-33.

[00461] [34] Lim TKY, Shi XQ, Johnson JM, Rone MB, Antel JP, David S, et al. Peripheral nerve injury induces persistent vascular dysfunction and endoneurial hypoxia, contributing to the genesis of neuropathic pain. Journal of Neuroscience. 2015;35:3346-59.

[00462] [35] Marques MJ, Santo Neto H. Imaging neuromuscular junctions by confocal fluorescence microscopy: individual endplates seen in whole muscles with vital intracellular staining of the nerve terminals. The Journal of Anatomy. 1998;192:425-30.

[00463] [36] Cherrick GR, Stein SW, Leevy CM, Davidson CS. Indocyanine green: observations on its physical properties, plasma decay, and hepatic extraction. Journal of Clinical Investigation. 1960;39:592.

[00464] [37] Marshall MV, Rasmussen JC, Tan IC, Aldrich MB, Adams KE, Wang X, et al. Near infrared fluorescence imaging in humans with indocyanine green: a review and update. Open surgical oncology journal (Online). 2010;2:12.

[00465] [38] Guillonneau B. Neurological and vascular preservation during laparoscopic radical prostatectomy. Progres en urologie: journal de l'Association francaise d'urologie et de la Societe francaise d'urologie. 2009;19:5180-2.

[00466] [39] Wang C, Wu C, Zhu J, Miller RH, Wang Y. Design, synthesis, and evaluation of coumarin-based molecular probes for imaging of myelination. Journal of medicinal chemistry. 2011;54:2331-40.

[00467] The examples set forth above are provided to give those of ordinary skill in the art a complete disclosure and description of how to make and use the embodiments of the compositions, systems and methods of the disclosure, and are not intended to limit the scope of what the inventors regard as their disclosure. Modifications of the above-described modes for carrying out the disclosure that are obvious to persons of skill in the art are intended to be within the scope of the following claims. All patents and publications mentioned in the specification are indicative of the levels of skill of those skilled in the art to which the disclosure pertains. All references cited in this disclosure are incorporated by reference to the same extent as if each reference had been incorporated by reference in its entirety individually.

[00468] All headings and section designations are used for clarity and reference purposes only and are not to be considered limiting in any way. For example, those of skill in the art will appreciate the usefulness of combining various aspects from different headings and sections as appropriate according to the spirit and scope of the invention described herein.

[00469] All references cited herein are hereby incorporated by reference herein in their entireties and for all purposes to the same extent as if each individual publication or patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety for all purposes.

[00470] Many modifications and variations of this application can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. The specific embodiments and examples described herein are offered by way of example only, and the application is to be limited only by the terms of the appended claims, along with the full scope of equivalents to which the claims are entitled.

Claims

1. A human neuron or nerve targeting molecule that specifically binds to a human neuron or nerve, or component of either, wherein said targeting molecule comprises a peptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac DLPDIIWDFNWETAGGC (HNP403; SEQ ID NO:6), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac-PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac-EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9), Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13), Ac SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21), PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23), PYYVVKKSS (HNP401-N-8; SEQ ID NO:24), SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), SGQVPWEEP (HNP401 C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122), SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), and SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124).

2. The human neuron or nerve targeting molecule of claim 1, wherein said targeting molecule comprises a peptide selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), Ac-SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac-SGQVPWEEPYYVVKKGGC (HNP401-C-2 with GGC linker; SEQ ID NO:11), Ac-QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7),

QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21), and SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25).

3. The human neuron or nerve targeting molecule of claim 1, wherein said targeting molecule comprises the peptide QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21).

4. The human neuron or nerve targeting molecule of any one of claims 1-3, further comprising a cargo selected from the group consisting of a drug, fluorescent moiety, and photosensitizing agent.

5. The human neuron or nerve targeting molecule of claim 4, wherein said targeting molecule comprises 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

6. The human neuron or nerve targeting molecule of claim 4, wherein said targeting molecule consists of 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

7. The human neuron or nerve targeting molecule of claim 4, wherein the cargo comprises the drug.

8. The human neuron or nerve targeting molecule of claim 7, wherein the drug is selected from the group consisting of: an antihistamine, a GABA receptor modulator, a neurotransmitter reuptake inhibitor, a local anesthetic, an anticholinergic, a sodium channel blocker, a calcium channel blocker, a thyrotropin-releasing hormone, a y-secretase inhibitor, an AMPA receptor agonist or antagonist, an NMDA receptor agonist or antagonist, an mGlu receptor agonist or antagonist, a growth factor, an antiemetic agent, a corticosteroid; a cytotoxic agent; an antioxidant, an iron chelator, a mitochondrial modulator, a sirtuin modulator, a nitric oxide (NO) and/or nitric oxide synthase (NOS) modulator, a potassium channel agonist or antagonist, a purigenic receptor agonist or antagonist, and combinations thereof.

9. The human neuron or nerve targeting molecule of claim 7, wherein the drug is selected from the group consisting of: benzocaine; carticaine; cinchocaine; cyclomethycaine; lidocaine; prilocaine; propxycaine; proparacaine; tetracaine; tocainide; and trimecaine; methotrexate; cyclophosphamide; thalidomide; paclitaxel; pemetrexed; pentostatin; pipobroman; pixantrone; plicamycin; platonin; procarbazine; raltitrexed; rebeccamycin; rubitecan; SN-38; salinosporamide A; satraplatin; streptozotocin; swainsonine; tariquidar; taxane; tegafur-uracil; temozolomide; testolactone; thioTEPA; tioguanine; topotecan; trabectedin; tretinoin; triplatin tetranitrate; tris(2- chloroethyl)amine; troxacitabine; uracil mustard; valrubicin; vinblastine; vincristine; vinorelbine; vorinostat; zosuquidar; carbamazepine; oxcarbazepine; phenytein; valproic acid; sodium valproate; cinnarizine; flunarizine; nimodipine; brain-derived neurotrophic factor (BDNF); ciliary neurotrophic factor (CNTF); glial cell-line derived neurotrophic factor (GDNF); neurotrophin-3; neurotrophin-4; fibroblast growth factor (FGF) receptor; insulin-like growth factor (IGF); and combinations thereof.

10. The human neuron or nerve targeting molecule of claim 4, wherein the cargo comprises the fluorescent moiety.

11. The human neuron or nerve targeting molecule of claim 10, wherein the fluorescent moiety is selected from the group consisting of: a fluorescent protein, a fluorescent peptide, a fluorescent dye, and combinations thereof.

12. The human neuron or nerve targeting molecule of claim 10, wherein the fluorescent moiety is selected from the group consisting of: a xanthene; a bimane; a coumarin; an aromatic amines; a benzofuran; a fluorescent cyanine; a carbazole; a dicyanomethylene pyrane; polymethine; oxabenzanthrane; pyrylium; carbostyl; perylene; acridone; quinacridone; rubrene; anthracene; coronene; phenanthrecene; pyrene; butadiene; stilbene; porphyrin; pthalocyanine; lanthanide metal chelate complexes; rare-earth metal chelate complexes; and derivatives thereof.

13. The human neuron or nerve targeting molecule of claim 10, wherein the fluorescent moiety is selected from the group consisting of: 5-carboxyfluorescein; fluorescein-5 isothiocyanate; 6-carboxyfluorescein; tetramethylrhodamine-6-isothiocyanate; 5 carboxytetramethylrhodamine; 5-carboxy rhodol derivatives; tetramethyl and tetraethyl rhodamine; diphenyldimethyl and diphenyldiethyl rhodamine; dinaphthyl rhodamine; rhodamine 101 sulfonyl chloride; Cy3, Cy3B, Cy3.5, Cy5, Cy5.5, Cy 7, indocyanine green, IR800CW, cyan fluorescent protein (CFP), EGFP, 6-FAM, FAM, fluorescein, 5,6-dicarboxyfluorescein, 5-(and 6)-sulfofluorescein, sulfonefluorescein, succinyl fluorescein, 5-(and 6)-carboxy SNARF-1, carboxyfluorescein sulfonate, carboxyfluorescein zwitterion, carboxyfluorescein quaternary ammonium, carboxyfluorescein phosphonate, carboxyfluorescein GABA, carboxyfluorescein cys-Cy5, 5'(6')-carboxyfluorescein, fluorescein glutathione, and combinations thereof.

14. The human neuron or nerve targeting molecule of claim 4, wherein the cargo comprises the photosensitizing agent.

15. The human neuron or nerve targeting molecule of claim 14, wherein the photosensitizing agent is selected from the group consisting of: a porphyrin, chlorin, and dye.

16. The human neuron or nerve targeting molecule of claim 14, wherein the photosensitizing agent selected from the group consisting of: porphyrin, protoporfin IX, purlytin, verteporfin, HPPH, temoporfin, methylene blue, photofrin, protofrin, hematoporphyrin, Talaporfin, benzopophyrin derivative monoacid, 5-aminileuvolinic acid, Lutetium texaphyrin, metallophthalocyanine, metallo-naphthocyaninesulfobenzo-porphyrazine, metallo naphthalocyanines, zinc tetrasulfophthalocyanine, bacteriochlorins, metallochlorins, chlorine derivative, Tetra(m-hydroxyphenyl)chlorin (mTHPC), pheophorbide, dibromofluorescein (DBF), IR700DX, naphthalocyanine, and porphyrin derivatives.

17. The human neuron or nerve targeting molecule of any one of claims 4-16, wherein the cargo is joined to the N-terminus of the peptide.

18. The human neuron or nerve targeting molecule of any one of claims 4-16, wherein the cargo is joined to the C-terminus of the peptide.

19. The human neuron or nerve targeting molecule of any one of claims 4-18, wherein the cargo is joined to the peptide via a linker.

20. The human neuron or nerve targeting molecule of claim 19, wherein the linker is a straight or branched-chain carbon linker, heterocyclic carbon linker, amino acid linker, lipophilic residue, peptide linker, peptide nucleic acid linker, hydrazone linker, SPDB disulfide, sulfo-SPDB, maleimidomethyl cyclohexane-1-carboxylate (MCC), aminohexanoic acid linker, polyether linker, or polyethylene glycol linker.

21. The human neuron or nerve targeting molecule of claim 19, wherein the linker is a peptide linker.

22. The human neuron or nerve targeting molecule of claim 21, wherein the peptide linker has less than 5 amino acids.

23. The human neuron or nerve targeting molecule of claim 21 or 22, wherein the peptide linker comprises a glycine linker.

24. The human neuron or nerve targeting molecule of claim 1, comprising: 5FAM QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104), Ac SGQVPWEEPYYVVKKSSGGC-5FAM (HNP401 with GGC linker; SEQ ID NO:105), Ac-

WEYHYVDLNWTSQHPQGGC-5FAM (HNP402 with GGC linker; SEQ ID NO:106), Ac DLPDIIWDFNWETAGGC-5FAM (HNP403 with GGC linker; SEQ ID NO:107), Ac QVPWEEPYYVVKKSSGGC-5FAM (HNP401-N-2 with GGC linker; SEQ ID NO:108), Ac PWEEPYYVVKKSSGGC-5FAM (HNP401-N-4 with GGC linker; SEQ ID NO:109), Ac EEPYYVVKKSSGGC-5FAM (HNP401-N-6 with GGC linker; SEQ ID NO:110), Ac PYYVVKKSSGGC-5FAM (HNP401-N-8 with GGC linker; SEQ ID NO:111), Ac SGQVPWEEPYYVVKKGGC-5FAM (HNP401-C-2 with GGC linker; SEQ ID NO:112), Ac SGQVPWEEPYYVVGGC-5FAM (HNP401-C-4 with GGC linker; SEQ ID NO:113), Ac SGQVPWEEPYYGGC-5FAM (HNP401-C-6 with GGC linker; SEQ ID NO:114), or Ac SGQVPWEEPGGC-5FAM (HNP401-C-8 with GGC linker; SEQ ID NO:115).

25. A multidomain neuron or nerve targeting molecule comprising two or more neuron or nerve targeting peptides, wherein the two or more neuron or nerve targeting peptides bind to a human neuron or nerve, or component of either, wherein the first peptide is selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9), Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac-SGQVPWEEPYYVVKKGGC (HNP401 C-2 with GGC linker; SEQ ID NO:11), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS (HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120),

SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122), SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), and 5FAM-QVPWEEPYYVVKKSSGG-NH2 (HNP401-N-2 with GG linker; SEQ ID NO:104).

26. The multidomain neuron or nerve targeting molecule of claim 25, wherein the second peptide is selected from the group consisting of: SGQVPWEEPYYVVKKSS (HNP 401; SEQ ID NO:1), WEYHYVDLNWTSQHPQ (HNP 402; SEQ ID NO:2), DLPDIIWDFNWETA (HNP 403; SEQ ID NO:3), DTHAHAKPRVPAFKSV (HNP 404; SEQ ID NO:16), Ac SGQVPWEEPYYVVKKSSGGC (HNP401 with GGC linker; SEQ ID NO:4), Ac WEYHYVDLNWTSQHPQGGC (HNP402 with GGC linker; SEQ ID NO:5), Ac DLPDIIWDFNWETAGGC (HNP403 with GGC linker; SEQ ID NO:6), Ac QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO:7), Ac PWEEPYYVVKKSSGGC (HNP401-N-4 with GGC linker; SEQ ID NO:8), Ac EEPYYVVKKSSGGC (HNP401-N-6 with GGC linker; SEQ ID NO:9), Ac-PYYVVKKSSGGC (HNP401-N-8 with GGC linker; SEQ ID NO:10), Ac-SGQVPWEEPYYVVKKGGC (HNP401 C-2 with GGC linker; SEQ ID NO:11), Ac-SGQVPWEEPYYVVGGC (HNP401-C-4 with GGC linker; SEQ ID NO:12), Ac-SGQVPWEEPYYGGC (HNP401-C-6 with GGC linker; SEQ ID NO:13), Ac-SGQVPWEEPGGC (HNP401-C-8 with GGC linker; SEQ ID NO:14), QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO:20); QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO:21); PWEEPYYVVKKSS (HNP401-N-4; SEQ ID NO:22); EEPYYVVKKSS (HNP401-N-6; SEQ ID NO:23); PYYVVKKSS (HNP401-N-8; SEQ ID NO:24); SGQVPWEEPYYVVKK (HNP401-C-2; SEQ ID NO:25), SGQVPWEEPYYVV (HNP401-C-4; SEQ ID NO:26), SGQVPWEEPYY (HNP401-C-6; SEQ ID NO:27), and SGQVPWEEP (HNP401-C-8; SEQ ID NO:28), PWEEPYYVVKKSSGG (HNP401-N-4 with GG linker; SEQ ID NO:118), EEPYYVVKKSSGG (HNP401-N-6 with GG linker; SEQ ID NO:119), PYYVVKKSSGG (HNP401-N-8 with GG linker; SEQ ID NO:120), SGQVPWEEPYYVVKKGG (HNP401-C-2; with GG linker; SEQ ID NO:121), SGQVPWEEPYYVVGG (HNP401-C-4 with GG linker; SEQ ID NO:122), SGQVPWEEPYYGG, (HNP401-C-6 with GG linker; SEQ ID NO:123), SGQVPWEEPGG (HNP401-C-8 with GG linker; SEQ ID NO:124), SHSSEFPRSWDMETN (HNP301; SEQ ID NO:29); SHSMLPSVLD (HNP303; SEQ ID NO:30); SHSTMKTLSL (HNP305; SEQ ID NO:31); VAPTKAPLHSPS (NP121; SEQ ID NO:32), NNLKTGTSAPTG (NP122; SEQ ID NO:33), HKTAQWPFIAFR (NP123; SEQ ID NO:34), RLTNAPAYQAPA (NP124; SEQ ID

NO:35), MQNPLNGKPGR (NP125; SEQ ID NO:36), THYSRSLTDGTR (NP126; SEQ ID NO:37), YPSPNRPPNLTN (NP127; SEQ ID NO:38), and NTQTLAKAPEHTG (NP117; SEQ ID NO:39); and/or the first peptide and the second peptide are the same or different.

27. The multidomain neuron or nerve targeting molecule of claim 25 or 26, wherein (1) the targeting molecule further comprises a cargo selected from the group consisting of a drug, fluorescent moiety, and photosensitizing agent; (2) the first peptide and second peptide are joined via a linker; and/or (3) the first peptide and second peptide are joined via the cargo.

28. A method of identifying a human neuron or nerve comprising contacting the human neuron or nerve with a targeting molecule according to any one of claims 4-6, 10-13 or 24, wherein the cargo comprises the fluorescent moiety.

29. A method of delivering a drug to a human neuron or nerve comprising contacting the human neuron or nerve with a human neuron or nerve targeting molecule according to any one of claims 4 or 7-9, wherein the cargo comprises the drug.

30. A method of delivering a photosensitizing agent to a human neuron or nerve comprising contacting the human neuron or nerve with a human neuron or nerve targeting molecule according to any one of claims 4 or 14-16, wherein the cargo comprises the photosensitizing agent.

31. The method of claim 30, further comprising exposing the human neuron or nerve to a light source that activates the photosensitizing agent, wherein the human neuron or nerve is ablated by the activated photosensitizing agent.

32. The method of any one of claims 28-31, wherein said human neuron or nerve targeting molecule is administered by systemic intravenous injection of a human subject.

33. The method of any one of claims 28-32, wherein said human neuron or nerve targeting molecule is administered prior to a surgical procedure.

34. The method of claim 33, wherein said surgical procedure is a cancer surgical procedure, optionally, wherein said surgical procedure is a prostate cancer surgical procedure.

35. The method of claim 33, wherein said surgical procedure is a trauma surgery, a reconstructive surgery, a head and neck surgery, a spinal surgery, a prostate surgery, or any combination thereof.

36. A pharmaceutical composition comprising: (a) the human neuron or nerve targeting molecule of any one of claims 1-24 or the multidomain neuron or nerve targeting molecule of any one of claims 25-27, and (b) a pharmaceutically acceptable excipient.

The Regents of the University of California Patent Attorneys for the Applicant/Nominated Person SPRUSON & FERGUSON

Figure 1

c2m1L4011 c2m1L401r

450nmols/2 hours post/6month old mice

m2np41 m2np41r

c2m3L401L c2m3L401r

S

m1np41 m1np41r

c2m2L401 c2m2L401r

SUBSTITUTE SHEET (RULE 26)

Figure 2

Quantitative comparison of HNP401 to NP41 in-vivo

1000

900

800

700

600

500 Nerva Muscle 400 300

200

100

0 HNP401 NP41

6.2 fold 6.7 fold

SUBSTITUTE SHEET (RULE 26)

Figure 3

Compliation of Various Nerve Peptides on Human Nerve Sections HNP-401 is the brightest but similar to NP-124 (high background staining)

Carboxyfluorescein HNP-401* HNP-402* HNP-404 Fresh sections

human nerve. Peptides treated on

10um dried sections at 300uM

in 0.5x HBSS except HNP402 which was in

NP-41 NP-124 NP-713 HNP-301 0.06xHBSS/water at room temperature for 20- 25 minutes in humidified

chamber Washed in PBS and imaged on Nikon Confocal with

488mm laser and 25x objective (4x4 Carboxyfluorescein D-np-41 D-np-713 tile), photoshop *Gain 10, scaled to 10 all others

gain=30 Otherwise scaled the same

Figure 4

Comparison of NP41, HNP401 and HNP41 binding to human nerve sections under identical capture setting and leveling

(100uM peptides from 52215)

Conc=100uM

NP41 HNP401 HNP404

SUBSTITUTE SHEET (RULE 26)

Figure 5

In vivo HNP-401 Rat sciatic nerve (81115)

Left Right

2 umols HNP401-FAM retro-orbital male rat 5 h circulation imaged in vivo

Standard conditions-12.2x 10s

SUBSTITUTE SHEET (RULE 26)

Figure 6

Comparison of white light and fluorescent images of HNP401 (9/15)/301(7/15 in RAT Prostate (HNP401 has higher background but better visibility of nerve!)

Retroorbital Vas route del 2 umoles HNP401 4 hour circulation 8x 5 S HNP401 White-light

2umols blander HNP301 Prostate imaged at préstate 12 x5 S expo Retroorbital

4 hours Vas

HNP301 def

SUBSTITUTE SHEET (RULE 26)

Figure 7

Nerve bundle

Blood

vesse

Bladder

12.2 X 1s exp 25x .5s exp

2 umols NP401-FAM male rat 5 h circulation imaged in vivo Standard conditions- 12.2 x 10s

SUBSTITUTE SHEET (RULE 26)

FIG. 8A FIG. 8B FIG. 8C

A B C

FAM-HNP407 FAM-HNP402 FAM-HNP403

E F

carboxyFAM FAM-NP41 nerve

muscle FIG. 8D FIG. 8E FIG. 8F

SUBSTITUTE SHEET (RULE 26)